Pyrimidine FGFR4 inhibitors

ABSTRACT

Provided herein are compounds of Formula I useful as FGFR4 inhibitors, as well as methods of use of the same.

BACKGROUND

Fibroblast growth factors (FGF) are a family of more than 20structurally related proteins with a variety of biological activities.Their main receptors, the fibroblast growth factor receptors (FGFR1,FGFR2, FGFR3 and FGFR4), are a family of receptor tyrosine kinases thatbind FGF and are involved in processes of cell proliferation anddifferentiation. Deregulation of FGFR signaling networks is implicatedin a number of pathophysiological conditions, including many types ofhuman cancers.

“Fibroblast Growth Factor Receptor 4” or “FGFR4” is known to regulateproliferation and antiapoptosis and is expressed or highly expressed inmany cancers. See, e.g., Dieci et al. 2013, Cancer Discovery, 0F1-0F16.Studies have shown that expression of FGFR4 is predictive of a moreaggressive phenotype of the cancer, and knockdown or reduction of FGFR4expression serves to reduce proliferation and promote apoptosis. See,e.g., Wesche et al. 2011, Biochem J 437:199-213.

For example, FGFR4 expression or overexpression is associated withcancer aggressiveness in gastric cancer (Ye et al. 2011, Cancer,5304-5313), prostate cancer (Xu et al. 2011, BMC Cancer, 11; 84),sarcoma such as rhabdomyosarcoma (Taylor V I et al. 2009, J Clin Invest,119(11):3395-3407), skin cancer such as melanoma (Streit et al. 2006,British J Cancer, 94:1879-1886), liver cancer such as cholangiocarcinoma(Sia et al. 2013, Gastroenterology 144:829-840) and hepatocellularcarcinoma (French et al. 2012, PLoS ONE 7(5): e367313; Miura et al.2012, BMC Cancer 12:56; Chiang et al. 2008, Cancer Res 68(16):6779-6788;Sawey et al. 2011, Cancer Cell 19:347-358), pancreatic cancer such aspancreatic intraepithelial neoplasia and pancreatic ductaladenocarcinoma (Motoda et al. 2011, Int'l J Oncol 38:133-143), lungcancer such as non-small-cell lung cancer (Fawdar et al. 2013, PNAS110(30):12426-12431), colorectal cancer (Pelaez-Garcia et al. 2013, PLoSONE 8(5): e63695; Barderas et al. 2012, J Proteomics 75:4647-4655), andovarian cancer (Zaid et al. 2013, Clin Cancer Res 19:809-820).

Clinical development of several FGFR inhibitors have confirmed theirutility as antitumor agents. Dieci et al. 2013, Cancer Discovery,0F1-0F16. However, new agents are needed that are useful to target FGFR,and FGFR4, in particular.

SUMMARY

A purpose of the present invention is to provide a compound of FormulaI:

wherein:

R³ is selected from the group consisting of: C₁₋₆alkyl,C₁₋₆alkoxyC₁₋₆alkyl, NR¹⁰R¹¹C₁₋₆alkyl, R¹⁰heterocyclylC₁₋₆alkyl,R¹⁰arylC₁₋₆alkyl, and R¹⁰heteroarylC₃₋₆alkyl, wherein R¹⁰ and

R¹¹ are each independently selected from the group consisting of:hydrogen and C₁₋₆alkyl;

E is selected from the group consisting of:

-   -   —NR¹³C(O)CR¹⁴═CHR¹⁵, and    -   —NR¹³C(O)C≡CR¹⁴,

wherein R¹³ is selected from the group consisting of: hydrogen andmethyl, and R¹⁴ and R¹⁵ are each independently selected from the groupconsisting of: hydrogen, methyl, fluoro and chloro;

R¹² is selected from the group consisting of: hydrogen, halo, C₁₋₆alkyl,C₁₋₆alkoxy, hydroxyC₁₋₆alkyl, hydroxyC₁₋₆alkoxy, C₁₋₆alkoxyC₁₋₆alkoxy,C₁₋₆alkoxyC₁₋₆alkyl, R⁵R⁶heterocyclyl, —C(O)heterocyclylR⁵R⁶,R⁵R⁶heterocyclylC₁₋₆alkyl, NR⁵R⁶, NR⁵R⁶C₁₋₆alkyl, —C(O)NR⁵R⁶, andNR⁵R⁶C₁₋₆alkyoxy, wherein R⁵ and R⁶ are each independently selected fromthe group consisting of hydrogen, C₁₋₆alkyl, hydroxyC₁₋₆alkyl,aminoC₁₋₆alkyl, —C(O)C₁₋₆alkyl and C₁₋₆alkylsulfonyl; and

R¹ is phenyl, wherein said phenyl is substituted 2, 3, or 4 times withindependently selected halo or C₁₋₆alkoxy,

or a pharmaceutically acceptable salt thereof.

In some embodiments, R³ is C₁₋₆alkyl.

In some embodiments, R³ is selected from the group consisting of:methyl, methoxyethyl, 4-pyridylmethyl, 3-pyridylmethyl, 2-pyridylmethyl,benzyl, N,N-dimethylaminopropyl, 3-methylisoxazol-5-yl-methyl, and4-methylpiperazin-1-yl-propyl.

In some embodiments, E is —NR¹³C(O)CH═CHR¹⁵ or —NR¹³C(O)CF═CH₂, whereinR¹³ and R¹⁵ are as defined above. In some embodiments, E is—NHC(O)CH═CH₂.

In some embodiments, R¹² is selected from the group consisting of:hydrogen, fluoro, chloro, methyl, methoxy, N,N-dimethylaminoethyl,piperazin-1-yl, 4-ethylpiperazin-1-yl, 4-ethylpiperazin-1-yl-methyl,1-methylpiperidine-4-yl, 1-ethylpiperidine-4-yl,N,N-dimethylaminomethyl, N,N-dimethylaminopropyl, piperidine-4-yl,morpholino, 3,5-dimethylpiperazin-1-yl,4-(methylsulfonyl)piperazin-1-yl, N,N-dimethylaminoethoxy,4-(2-hydroxyethyl)piperazin-1-yl, hydroxyethoxy, methoxyethoxy,hydroxymethyl, methoxymethyl, 2-methoxypropyl, 2-hydroxypropyl,2-aminopropyl, 4-methylpiperazin-1-yl-carbonyl,4-ethylpiperazin-1-yl-carbonyl, 4-[2-propyl]piperazin-1-yl,4-acetylpiperazin-1-yl, N-methyl-N-hydroxyethyl-amino, N,N-dimethylamido, and 4-(2-aminoethyl)piperazin-1-yl.

In some embodiments, R¹² is selected from the group consisting of:hydrogen, C₁₋₆alkyl, hydroxyC₁₋₆alkyl, R⁵R⁶heterocyclyl,R⁵R⁶heterocyclylC₁₋₆alkyl, —C(O)NR⁵R⁶, NR⁵R⁶C₁₋₆alkyl, NR⁵R⁶C₁₋₆alkyoxy,C₁₋₆alkoxy, and C₁₋₆alkoxyC₁₋₆alkyl, wherein R⁵ and R⁶ are eachindependently selected from the group consisting of: hydrogen,C₁₋₆alkyl, hydroxyC₁₋₆alkyl, —C(O)C₁₋₆alkyl and C₁₋₆alkylsulfonyl.

In some embodiments, R¹² is R⁵R⁶heterocyclyl, wherein R⁵ and R⁶ are asdefined above.

In some embodiments, R⁵R⁶heterocyclyl is R⁵R⁶piperazinyl, wherein R⁵ andR⁶ are as defined above.

In some embodiments, R¹² is 4-ethylpiperazin-1-yl.

In some embodiments, R¹² is not hydrogen.

In some embodiments, R¹ is 2,6-dichloro-3,5-dimethoxyphenyl.

In some embodiments, the compound is a compound of Formula I(a):

wherein R³, E, R¹² and R¹ are as defined above,

or a pharmaceutically acceptable salt thereof.

A further purpose is a pharmaceutical composition comprising a compoundor salt as described herein and a pharmaceutically acceptable carrier.In some embodiments, the composition is formulated for oral orparenteral administration.

A further purpose is a method of treating hepatocellular carcinoma in asubject in need thereof comprising administering to said subject atreatment effective amount of a compound or salt or composition asdescribed herein. In some embodiments, hepatocellular carcinoma hasaltered FGFR4 and/or FGF19 status (e.g., increased expression of FGFR4and/or FGF19).

A further purpose is a method of treating hepatocellular carcinoma in asubject in need thereof, comprising: detecting an altered FGFR4 and/orFGF19 status (e.g., increased expression of FGFR4 and/or FGF19) in abiological sample containing cells of said hepatocellular carcinoma, andif said hepatocellular carcinoma has said altered FGFR4 and/or FGF19status, administering a compound or composition described herein to saidsubject in a treatment-effective amount.

A further purpose is the use of a compound or salt or a composition asdescribed herein in a method of treatment of hepatocellular carcinoma.

A further purpose is the use of a compound or salt described herein inthe preparation of a medicament for the treatment of hepatocellularcarcinoma.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 presents the results of in vivo efficacy testing inhepatocellular carcinoma model using HUH7 cells. Compound 108 (25 mg/kgor 37.5 mg/kg) or Vehicle control was administered via intraperitonealinjection, and tumor volume was measured twice weekly over the course of15 days.

FIG. 2 presents the results of in vivo efficacy testing inhepatocellular carcinoma model using HEP3B cells. Compound 108 (12.5mg/kg, 25 mg/kg or 37.5 mg/kg) or Vehicle control was administered viaintraperitoneal injection, and tumor volume was measured twice weeklyover the course of 15 days.

FIG. 3 presents the results of in vivo efficacy testing inhepatocellular carcinoma model using JHH7 cells. Compound 108 (12.5mg/kg, 25 mg/kg or 37.5 mg/kg) or Vehicle control was administered viaintraperitoneal injection, and tumor volume was measured twice weeklyover the course of 15 days.

FIG. 4 presents the results of comparative in vivo efficacy testing inhepatocellular carcinoma model using HEP3B cells. Compound 108 (25mg/kg, 37.5 mg/kg or 50 mg/kg) was administered twice daily viaintraperitoneal injection, or BGJ398 (30 mg/kg or 60 mg/kg) wasadministered orally twice daily.

DETAILED DESCRIPTION OF EMBODIMENTS

Provided herein are compounds useful as FGFR4 inhibitors. In someembodiments, the compounds are selective FGFR4 inhibitors in that theyhave a greater binding affinity and/or inhibitory effect of FGFR4 ascompared to that of FGFR1 and/or FGFR2 and/or FGFR3 (e.g., by 10-fold,100-fold, or 1000-fold greater or more).

A. Definitions

Compounds useful as active agents in accordance with the presentdisclosure include those described generally above and below, and arefurther illustrated by the embodiments, sub-embodiments, and speciesdisclosed herein. As used herein, the following definitions shall applyunless otherwise indicated.

As described herein, compounds of the invention may optionally besubstituted with one or more substituents, such as those illustratedgenerally herein, or as exemplified by particular classes, subclasses,and species of the invention. In general, the term “substituted” refersto the replacement of hydrogen in a given structure with a specifiedsubstituent. Unless otherwise indicated, a substituted group may have asubstituent at each substitutable position of the group, and when morethan one position in any given structure may be substituted with morethan one substituent selected from a specified group, the substituentmay be either the same or different at every position. Combinations ofsubstituents envisioned by this invention are preferably those thatresult in the formation of stable compounds. “Stable” as used hereinrefers to chemically feasible compound is one that is not substantiallyaltered when kept at a temperature of 40° C. or less, in the absence ofmoisture or other chemically reactive conditions, for at least a week.

As would be understood by those of skill in the art, as used herein “H”is hydrogen, “C” is carbon, “N” is nitrogen, “S” is sulfur, and “O” isoxygen.

“Alkyl” or “alkyl group,” as used herein, means a straight-chain (i.e.,unbranched), or branched hydrocarbon chain that is completely saturated.In some embodiments, the alkyl has 1, 2, 3, 4, 5 or 6 carbon atoms. Incertain embodiments, alkyl groups contain 1-6 carbon atoms (C₁₋₆alkyl).In certain embodiments, alkyl groups contain 1-4 carbon atoms(C₁₋₄alkyl). In certain embodiments, alkyl groups contain 1-3 carbonatoms (C₁₋₃alkyl). In still other embodiments, alkyl groups contain 2-3carbon atoms (C₂₋₃alkyl), and in yet other embodiments alkyl groupscontain 1-2 carbon atoms (C₁₋₂alkyl).

“Alkenyl” or “alkenyl group,” as used herein, refers to a straight-chain(i.e., unbranched), or branched hydrocarbon chain that has one or moredouble bonds. In some embodiments, the alkenyl has 2, 3, 4, 5 or 6carbon atoms. In certain embodiments, alkenyl groups contain 2-8 carbonatoms (C₂₋₈alkyl). In certain embodiments, alkenyl groups contain 2-6carbon atoms (C₂₋₆alkyl). In still other embodiments, alkenyl groupscontain 3-4 carbon atoms (C₃₋₄alkyl), and in yet other embodimentsalkenyl groups contain 2-3 carbon atoms (C₂₋₃alkyl). According toanother aspect, the term alkenyl refers to a straight chain hydrocarbonhaving two double bonds, also referred to as “diene.” Non-limitingexamples of exemplary alkenyl groups include —CH═CH₂, —CH₂CH═CH₂,—CH═CHCH₃, —CH₂CH₂CH═CH₂, —CH₂CH═CHCH₃, —CH═CHCH₂CH₃, and —CH═CHCH═CH₂.

“Alkynyl” or “alkynyl group” as used herein refers to a straight-chain(i.e., unbranched), or branched hydrocarbon chain that has one or moretriple bonds. In some embodiments, the alkynyl has 2, 3, 4, 5 or 6carbon atoms. In certain embodiments, alkynyl groups contain 2-8 carbonatoms (C₂₋₈alkynyl). In certain embodiments, alkynyl groups contain 2-6carbon atoms (C₂₋₆alkynyl). In still other embodiments, alkynyl groupscontain 3-4 carbon atoms (C₃₋₄alkynyl), and in yet other embodimentsalkynyl groups contain 2-3 carbon atoms (C₂₋₃alkynyl).

“Ar” or “aryl” refer to an aromatic carbocyclic moiety having one ormore closed rings. Examples include, without limitation, phenyl,naphthyl, anthracenyl, phenanthracenyl, biphenyl, and pyrenyl.

“Halo” refers to chloro (Cl), fluoro (F), bromo (Br) or iodo (I).

“Haloalkyl” refers to one or more halo groups appended to the parentmolecular moiety through an alkyl group. Examples include, but are notlimited to, chloromethyl, fluoromethyl, trifluoromethyl, etc.

“Heteroaryl” refers to a cyclic moiety having one or more closed rings,with one or more heteroatoms (oxygen, nitrogen or sulfur) in at leastone of the rings, wherein at least one of the rings is aromatic, andwherein the ring or rings may independently be fused, and/or bridged.Examples include, without limitation, quinolinyl, isoquinolinyl,indolyl, furyl, thienyl, pyrazolyl, quinoxalinyl, pyrrolyl, indazolyl,thieno[2,3-c]pyrazolyl, benzofuryl, pyrazolo[1,5-a]pyridyl,thiophenylpyrazolyl, benzothienyl, benzothiazolyl, thiazolyl,2-phenylthiazolyl, and isoxazolyl.

“—OR” or “oxy” refers to an R group appended to the parent molecularmoiety through an oxygen atom, wherein R is H, alkyl, alkenyl, alkynyl,and the like.

“Alkoxy” refers to an alkyl group, as herein defined, attached to theprincipal carbon chain through an oxygen (“alkoxy”) atom. Representativeexamples of “alkoxy” include, but are not limited to, methoxy, ethoxy,propoxy, phenoxy, 2-propoxy, butoxy, tert-butoxy, pentyloxy, hexyloxyand the like.

“Hydroxy” refers to an —OH group.

“Carbonyl” is a group having a carbon atom double-bonded to an oxygenatom (C═O), often depicted in chemical formulae as C(O).

An “acetyl” is a group —C(O)CH₃.

An “amine” or “amino” refers to a group —NH₂, wherein none, one or twoof the hydrogens may replaced by a suitable substituent as describedherein, such as alkyl, alkenyl, alkynyl, and the like.

An “amide” or “amido” refers to a group having a carbonyl bonded to anitrogen atom, such as —C(O)NH₂, wherein none, one or two of thehydrogens may replaced by a suitable substituent as described herein,such as alkyl, alkenyl, alkynyl, and the like.

“—SR” refers to an R group appended to the parent molecular moietythrough a sulfur atom, wherein R is alkyl, alkenyl, alkynyl, aryl,cycloalkyl, heterocyclo, or heteroaryl. Representative examples of “—SR”include, but are not limited to, ethanethiol, 3-methyl-1-butanethiol,phenylthiol and the like.

“Cycloalkyl” as used herein, refers to a saturated cyclic hydrocarbongroup containing from 3 to 8 carbons or more. Representative examples ofcycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

“Cycloalkenyl” as used herein, refers to an unsaturated cyclichydrocarbon group containing from 3 to 8 carbons or more and having oneor more double bonds.

“Cycloalkynyl” as used herein, refers to an unsaturated cyclichydrocarbon group containing from 3 to 8 carbons or more and having oneor more triple bonds.

“Electrophile” as used herein refers to a group having reduced electrondensity, typically comprising a carbon atom that is directly bonded to amore electronegative atom, such as an oxygen, nitrogen or halo.Exemplary electrophiles include, but are not limited to, diazomethane,trimethylsilyldiazomethane, alkyl halides, such as for example methyliodide, benzyl bromide and the like, alkyl triflates, such as forexample methyl triflate and the like, alkyl sulfonates, such as forexample ethyl toluenesulfonate, butyl methanesulfonate and the like,acyl halides, such as for example acetyl chloride, benzoyl bromide andthe like, acid anhydrides, such as for example acetic anhydride,succinic anhydride, maleic anhydride and the like, isocyanates, such asfor example methyl isocyanate, phenyl isocyanate and the like,isothiocyanates, such as for example methyl isothiocyanate, phenylisothiocyanate and the like, chloroformates, such as for example methylchloroformate, ethyl chloroformate, benzyl chloroformate and the like,sulfonyl halides, such as for example methanesulfonyl chloride,methanesulfonyl fluoride, p-toluenesulfonyl chloride and the like, silylhalides, such as for example trimethylsilyl chloride,tert-butyldimethylsilyl chloride and the like, phosphoryl halides suchas for example dimethyl chlorophosphate and the like, epoxides such asfor example 2-methyloxirane, aziridines such as for example2-methylaziridine, alpha-haloketone such as for example1-chloro-2-propanone, alpha-beta-unsaturated carbonyl compounds such asfor example acrolein, methyl vinyl ketone, cinnamaldehyde,N,N-dimethylacrylamide and the like, andgamma-halo-alpha-beta-unsaturated carbonyl compounds such as for example(E)-6-chlorohex-4-en-3-one. In some embodiments, electrophiles arealpha-haloketones, isothiocyanates, epoxides, aziridines, sulfonylhalides, or alpha-beta-unsaturated carbonyls.

In some embodiments, the electrophile is a Michael acceptor. As known inthe art, a “Michael acceptor” is an alkene or alkyne of the form

; wherein Z comprises an electron withdrawing group, including, but notlimited to, CHO, COR, COOR, CONRR′, CONROR′, CN, NO₂, SOR, SO₂R. R maybe H, alkyl, or aryl; wherein R′ is alkyl, alkenyl, alkoxy or aryl. Inanother embodiment, azodicarboxamides and quinones are Michaelacceptors. See, Santos, M. M. M. and Moreira, R., Mini-Reviews inMedicinal Chemistry, 7:1040-1050, 2007. An example of the MichaelReaction is depicted in the scheme below:

wherein electron withdrawing groups Z, Z′ and Z″ are as described above.In some embodiments, the Michael acceptors are alpha-beta-unsaturatedcarbonyl compounds including, but not limited to, alpha-beta-unsaturatedamides, alpha-beta-unsaturated ketones, alpha-beta-unsaturated esters,conjugated alkynyl carbonyls and alpha-beta-unsaturated nitriles.

“Alpha-beta-unsaturated amide” or “unsaturated amide” as used hereinrefers to an amide comprising an alkene or alkyne bonded directly to theamide carbonyl group and is represented by the structure

wherein R′ is hydrogen or alkyl.

“Heteroatom” refers to O, S or N.

“Heterocycle” or “heterocyclyl” as used herein, means a monocyclicheterocycle, a bicyclic heterocycle, or a tricyclic heterocyclecontaining at least one heteroatom in the ring.

The monocyclic heterocycle is a 3-, 4-, 5-, 6-, 7, or 8-membered ringcontaining at least one heteroatom independently selected from the groupconsisting of O, N, and S. In some embodiments, the heterocycle is a 3-or 4-membered ring containing one heteroatom selected from the groupconsisting of O, N and S. In some embodiments, the heterocycle is a5-membered ring containing zero or one double bond and one, two or threeheteroatoms selected from the group consisting of O, N and S. In someembodiments, the heterocycle is a 6-, 7-, or 8-membered ring containingzero, one or two double bonds and one, two or three heteroatoms selectedfrom the group consisting of O, N and S. Representative examples ofmonocyclic heterocycle include, but are not limited to, azetidinyl,azepanyl, aziridinyl, diazepanyl, 1,3-dioxanyl, 1,3-dioxolanyl,dihydropyranyl (including 3,4-dihydro-2H-pyran-6-yl), 1,3-dithiolanyl,1,3-dithianyl, imidazolinyl, imidazolidinyl, isothiazolinyl,isothiazolidinyl, isoxazolinyl, isoxazolidinyl, morpholinyl,oxadiazolinyl, oxadiazolidinyl, oxazolinyl, oxazolidinyl, piperazinyl,piperidinyl, pyranyl, pyrazolinyl, pyrazolidinyl, pyrrolinyl,pyrrolidinyl, tetrahydrofuranyl, tetrahydropyranyl (includingtetrahydro-2H-pyran-4-yl), tetrahydrothienyl, thiadiazolinyl,thiadiazolidinyl, thiazolinyl, thiazolidinyl, thiomorpholinyl,1,1-dioxidothiomorpholinyl (thiomorpholine sulfone), thiopyranyl, andtrithianyl.

The bicyclic heterocycles of the present invention may be exemplified bya monocyclic heterocycle fused to an aryl group, or a monocyclicheterocycle fused to a monocyclic cycloalkyl, or a monocyclicheterocycle fused to a monocyclic cycloalkenyl, or a monocyclicheterocycle fused to a monocyclic heterocycle. Representative examplesof bicyclic heterocycles include, but are not limited to,3,4-dihydro-2H-pyranyl, 1,3-benzodioxolyl, 1,3-benzodithiolyl,2,3-dihydro-1,4-benzodioxinyl, 2,3-dihydro-1-benzofuranyl,2,3-dihydro-1-benzothienyl, 2,3-dihydro-1H-indolyl,3,4-dihydroquinolin-2(1H)-one and 1,2,3,4-tetrahydroquinolinyl.

The tricyclic heterocycle is a bicyclic heterocycle fused to an arylgroup, or a bicyclic heterocycle fused to a monocyclic cycloalkyl, or abicyclic heterocycle fused to a monocyclic cycloalkenyl, or a bicyclicheterocycle fused to a monocyclic heterocycle. Representative examplesof tricyclic heterocycles include, but are not limited to,2,3,4,4a,9,9a-hexahydro-1H-carbazolyl,5a,6,7,8,9,9a-hexahydrodibenzo[b,d]furanyl, and5a,6,7,8,9,9a-hexahydrodibenzo[b, d]thienyl.

In the above heteroaryl and heterocycles the nitrogen or sulfur atomscan be optionally oxidized to various oxidation states. In a specificexample, the group S(O)₀₋₂ refers to —S-(sulfide), —S(O)— (sulfoxide),and —SO₂— (sulfone) respectively. For convenience, nitrogens,particularly but not exclusively, those defined as annular aromaticnitrogens, are meant to include those corresponding N-oxide forms.

“Pharmaceutically acceptable salt” as used herein refer to acid additionsalts or base addition salts of the compounds in the present disclosure.A pharmaceutically acceptable salt is any salt which retains theactivity of the parent compound and does not impart any undulydeleterious or undesirable effect on a subject to whom it isadministered and in the context in which it is administered.Pharmaceutically acceptable salts include, but are not limited to, metalcomplexes and salts of both inorganic and carboxylic acids.Pharmaceutically acceptable salts also include metal salts such asaluminum, calcium, iron, magnesium, manganese and complex salts. Inaddition, pharmaceutically acceptable salts include, but are not limitedto, acid salts such as acetic, aspartic, alkylsulfonic, arylsulfonic,axetil, benzenesulfonic, benzoic, bicarbonic, bisulfuric, bitartaric,butyric, calcium edetate, camsylic, carbonic, chlorobenzoic, citric,edetic, edisylic, estolic, esyl, esylic, formic, fumaric, gluceptic,gluconic, glutamic, glycolic, glycolylarsanilic, hexamic,hexylresorcinoic, hydrabamic, hydrobromic, hydrochloric, hydroiodic,hydroxynaphthoic, isethionic, lactic, lactobionic, maleic, malic,malonic, mandelic, methanesulfonic, methylnitric, methylsulfuric, mucic,muconic, napsylic, nitric, oxalic, p-nitromethanesulfonic, pamoic,pantothenic, phosphoric, monohydrogen phosphoric, dihydrogen phosphoric,phthalic, polygalactouronic, propionic, salicylic, stearic, succinic,sulfamic, sulfanlic, sulfonic, sulfuric, tannic, tartaric, teoclic,toluenesulfonic, and the like. Pharmaceutically acceptable salts may bederived from amino acids including, but not limited to, cysteine.Methods for producing compounds as salts are known to those of skill inthe art (see, e.g., Stahl et al., Handbook of Pharmaceutical Salts:Properties, Selection, and Use, Wiley-VCH; Verlag Helvetica ChimicaActa, Zurich, 2002; Berge et al., J. Pharm. Sci. 66: 1, 1977).

Unless indicated otherwise, nomenclature used to describe chemicalgroups or moieties as used herein follow the convention where, readingthe name from left to right, the point of attachment to the rest of themolecule is at the right-hand side of the name. For example, the group“arylC₁₋₆alkyl” is attached to the rest of the molecule at the alkylend.

Unless indicated otherwise, where a chemical group is described by itschemical formula, including a terminal bond moiety indicated by “-,” itwill be understood that the attachment is read from left to right. Forexample, —C(O)C₁₋₆alkyl is attached to the rest of the molecule at thecarbonyl end.

Unless otherwise stated, structures depicted herein are also meant toinclude all enantiomeric, diastereomeric, and geometric (orconformational) forms of the structure; for example, the R and Sconfigurations for each asymmetric center, (Z) and (E) double bondisomers, and (Z) and (E) conformational isomers. Therefore, singlestereochemical isomers as well as enantiomeric, diastereomeric, andgeometric (or conformational) mixtures of the present compounds arewithin the scope of the invention. Unless otherwise stated, alltautomeric forms of the compounds of the invention are within the scopeof the invention. In addition, unless otherwise stated, all rotamerforms of the compounds of the invention are within the scope of theinvention. Unless otherwise stated, structures depicted herein are alsomeant to include compounds that differ only in the presence of one ormore isotopically enriched atoms. For example, compounds having thepresent structures except for the replacement of hydrogen by deuteriumor tritium, or the replacement of a carbon by a ¹³C- or ¹⁴C-enrichedcarbon are within the scope of this invention. Such compounds areuseful, for example, as analytical tools or probes in biological assays.

“Isomers” refer to compounds having the same number and kind of atomsand hence the same molecular weight, but differing with respect to thearrangement or configuration of the atoms. It will be understood,however, that some isomers or racemates or others mixtures of isomersmay exhibit more activity than others. “Stereoisomers” refer to isomersthat differ only in the arrangement of the atoms in space.“Diastereoisomers” refer to stereoisomers that are not mirror images ofeach other. “Enantiomers” refers to stereoisomers that arenon-superimposable mirror images of one another.

In some embodiments, enantiomeric compounds taught herein may be“enantiomerically pure” isomers that comprise substantially a singleenantiomer, for example, greater than or equal to 90%, 92%, 95%, 98%, or99%, or equal to 100% of a single enantiomer.

In some embodiments, enantiomeric compounds taught herein may bestereomerically pure. “Stereomerically pure” as used herein means acompound or composition thereof that comprises one stereoisomer of acompound and is substantially free of other stereoisomers of thatcompound. For example, a stereomerically pure composition of a compoundhaving one chiral center will be substantially free of the oppositeenantiomer of the compound. A stereomerically pure composition of acompound having two chiral centers will be substantially free ofdiastereomers, and substantially free of the opposite enantiomer, of thecompound. A typical stereomerically pure compound comprises greater thanabout 80% by weight of one stereoisomer of the compound and less thanabout 20% by weight of the other stereoisomers of the compound, morepreferably greater than about 90% by weight of one stereoisomer of thecompound and less than about 10% by weight of the other stereoisomers ofthe compound, even more preferably greater than about 95% by weight ofone stereoisomer of the compound and less than about 5% by weight of theother stereoisomers of the compound, and most preferably greater thanabout 97% by weight of one stereoisomer of the compound and less thanabout 3% by weight of the other stereoisomers of the compound. See,e.g., U.S. Pat. No. 7,189,715.

“R” and “S” as terms describing isomers are descriptors of thestereochemical configuration at an asymmetrically substituted carbonatom. The designation of an asymmetrically substituted carbon atom as“R” or “S” is done by application of the Cahn-Ingold-Prelog priorityrules, as are well known to those skilled in the art, and described inthe International Union of Pure and Applied Chemistry (IUPAC) Rules forthe Nomenclature of Organic Chemistry. Section E, Stereochemistry.

“Enantiomeric excess” (ee) of an enantiomer is [(the mole fraction ofthe major enantiomer) minus (the mole fraction of the minorenantiomer)]×100.

B. Compounds

Provided herein as active agents according to some embodiments is acompound of Formula I:

wherein:

R³ is selected from the group consisting of: C₁₋₆alkyl,C₁₋₆alkoxyC₁₋₆alkyl, NR¹⁰R¹¹C₁₋₆alkyl, R¹⁰heterocyclylC₁₋₆alkyl,R¹⁰arylC₁₋₆alkyl, and R¹⁰heteroarylC₁₋₆alkyl, wherein R¹⁰ and R¹¹ areeach independently selected from the group consisting of: hydrogen andC₁₋₆alkyl;

E is selected from the group consisting of:

-   -   —N¹³C(O)CR¹⁴═CHR¹⁵, and    -   —NR¹³C(O)C≡CR¹⁴,

wherein R¹³ is selected from the group consisting of: hydrogen andmethyl, and R¹⁴ and R¹⁵ are each independently selected from the groupconsisting of: hydrogen, methyl, fluoro and chloro;

R¹² is selected from the group consisting of: hydrogen, halo, C₁₋₆alkyl,C₁₋₆alkoxy, hydroxyC₁₋₆alkyl, hydroxyC₁₋₆alkoxy, C₁₋₆alkoxyC₁₋₆alkoxy,C₁₋₆alkoxyC₁₋₆alkyl, R⁵R⁶heterocyclyl, —C(O)heterocyclylR⁵R⁶,R⁵R⁶heterocyclylC₁₋₆alkyl, NR⁵R⁶, NR⁵R⁶C₁₋₆alkyl, —C(O)NR⁵R⁶, andNR⁵R⁶C₁₋₆alkyoxy, wherein R⁵ and R⁶ are each independently selected fromthe group consisting of hydrogen, C₁₋₆alkyl, hydroxyC₁₋₆alkyl,aminoC₁₋₆alkyl, —C(O)C₁₋₆alkyl and C₁₋₆alkylsulfonyl; and

R¹ is phenyl, wherein said phenyl is substituted 2, 3, or 4 times withindependently selected halo or C₁₋₆alkoxy,

or a pharmaceutically acceptable salt thereof.

In some embodiments, R³ is C₁₋₆alkyl.

In some embodiments, R³ is selected from the group consisting of:methyl, methoxyethyl, 4-pyridylmethyl, 3-pyridylmethyl, 2-pyridylmethyl,benzyl, N,N-dimethylaminopropyl, 3-methylisoxazol-5-yl-methyl, and4-methylpiperazin-1-yl-propyl.

In some embodiments, E is —NR¹³C(O)CH═CHR¹⁵ or —NR¹³C(O)CF═CH₂, whereinR¹³ and R¹⁵ are as defined above. In some embodiments, E is—NHC(O)CH═CH₂.

In some embodiments, R¹² is selected from the group consisting of:hydrogen, fluoro, chloro, methyl, methoxy, N,N-dimethylaminoethyl,piperazin-1-yl, 4-ethylpiperazin-1-yl, 4-ethylpiperazin-1-yl-methyl,1-methylpiperidine-4-yl, 1-ethylpiperidine-4-yl,N,N-dimethylaminomethyl, N,N-dimethylaminopropyl, piperidine-4-yl,morpholino, 3,5-dimethylpiperazin-1-yl,4-(methylsulfonyl)piperazin-1-yl, N,N-dimethylaminoethoxy,4-(2-hydroxyethyl)piperazin-1-yl, hydroxyethoxy, methoxyethoxy,hydroxymethyl, methoxymethyl, 2-methoxypropyl, 2-hydroxypropyl,2-aminopropyl, 4-methylpiperazin-1-yl-carbonyl,4-ethylpiperazin-1-yl-carbonyl, 4-[2-propyl]piperazin-1-yl,4-acetylpiperazin-1-yl, N-methyl-N-hydroxyethyl-amino,N,N-dimethylamido, and 4-(2-aminoethyl)piperazin-1-yl.

In some embodiments, R¹² is selected from the group consisting of:hydrogen, C₁₋₆alkyl, hydroxyC₁₋₆alkyl, R⁵R⁶heterocyclyl,R⁵R⁶heterocyclylC₁₋₆alkyl, —C(O)NR⁵R⁶, NR⁵R⁶C₁₋₆alkyl, NR⁵R⁶C₁₋₆alkyoxy,C₁₋₆alkoxy, and C₁₋₆alkoxyC₁₋₆alkyl, wherein R⁵ and R⁶ are eachindependently selected from the group consisting of: hydrogen,C₁₋₆alkyl, hydroxyC₁₋₆alkyl, —C(O)C₁₋₆alkyl and C₁₋₆alkylsulfonyl.

In some embodiments, R¹² is R⁵R⁶heterocyclyl, wherein R⁵ and R⁶ are asdefined above.

In some embodiments, R⁵R⁶heterocyclyl is R⁵R⁶piperazinyl, wherein R⁵ andR⁶ are as defined above.

In some embodiments, R¹² is 4-ethylpiperazin-1-yl.

In some embodiments, R¹² is not hydrogen.

In some embodiments, R¹ is 2,6-dichloro-3,5-dimethoxyphenyl.

In some embodiments, the compound is a compound of Formula I(a):

wherein R³, E, R¹² and R¹ are as defined above,

or a pharmaceutically acceptable salt thereof.

C. Pharmaceutical Formulations

Active agents of the present invention can be combined with apharmaceutically acceptable carrier to provide pharmaceuticalformulations thereof. The particular choice of carrier and formulationwill depend upon the particular route of administration for which thecomposition is intended.

“Pharmaceutically acceptable carrier” as used herein refers to anontoxic carrier, adjuvant, or vehicle that does not destroy thepharmacological activity of the compound with which it is formulated.Pharmaceutically acceptable carriers, adjuvants or vehicles that may beused in the compositions of this invention include, but are not limitedto, sorbic acid, potassium sorbate, partial glyceride mixtures ofsaturated vegetable fatty acids, water, salts or electrolytes, disodiumhydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zincsalts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone,cellulose-based substances, polyethylene glycol, sodiumcarboxymethylcellulose, polyacrylates, waxes, polyethylene glycol andwool fat.

The compositions of the present invention may be suitable forparenteral, oral, inhalation spray, topical, rectal, nasal, buccal,vaginal or implanted reservoir administration, etc. In some embodiments,the formulation comprise ingredients that are from natural ornon-natural sources.

In some embodiments, the formulation or carrier may be provided in asterile form. Non-limiting examples of a sterile carrier includeendotoxin-free water or pyrogen-free water.

The term “parenteral” as used herein includes subcutaneous, intravenous,intramuscular, intra-articular, intra-synovial, intrasternal,intrathecal, intrahepatic, intralesional and intracranial injection orinfusion techniques. In particular embodiments, the compounds areadministered intravenously, orally, subcutaneously, or via intramuscularadministration. Sterile injectable forms of the compositions of thisinvention may be aqueous or oleaginous suspension. These suspensions maybe formulated according to techniques known in the art using suitabledispersing or wetting agents and suspending agents. The sterileinjectable preparation may also be a sterile injectable solution orsuspension in a nontoxic parenterally acceptable diluent or solvent.Among the acceptable vehicles and solvents that may be employed arewater, Ringer's solution and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium.

For this purpose, any bland fixed oil may be employed includingsynthetic mono- or di-glycerides. Fatty acids and their glyceridederivatives are useful in the preparation of injectables, as are naturalpharmaceutically acceptable oils, such as olive oil or castor oil,especially in their polyoxyethylated versions. These oil solutions orsuspensions may also contain a long-chain alcohol diluent or dispersant,such as carboxymethyl cellulose or similar dispersing agents that arecommonly used in the formulation of pharmaceutically acceptable dosageforms including emulsions and suspensions. Other commonly usedsurfactants, such as Tweens, Spans and other emulsifying agents that arecommonly used in the manufacture of pharmaceutically acceptable solid,liquid, or other dosage forms may also be used for the purposes offormulation.

For oral administration, a compound or salt may be provided in anacceptable oral dosage form, including, but not limited to, capsules,tablets, aqueous suspensions or solutions. In the case of tablets fororal use, carriers commonly used include lactose and corn starch.Lubricating agents, such as magnesium stearate, may also be added. Fororal administration in a capsule form, useful diluents include lactoseand dried cornstarch. When aqueous suspensions are required for oraluse, the active ingredient may be combined with emulsifying andsuspending agents. If desired, certain sweetening, flavoring or coloringagents may also be added. In addition preservatives may also be added.Suitable examples of pharmaceutically acceptable preservatives include,but are not limited to, various antibacterial and antifungal agents suchas solvents, for example ethanol, propylene glycol, benzyl alcohol,chlorobutanol, quaternary ammonium salts, and parabens (such as methylparaben, ethyl paraben, propyl paraben, etc.).

D. Subjects and Methods of Use

Active agents of the present invention may be used to treathepatocellular carcinoma.

“Treatment,” “treat,” and “treating” refer to reversing, alleviating,delaying the onset of, inhibiting the progress of, or otherwiseameliorating a disease or disorder as described herein. In someembodiments, treatment may be administered after one or more symptomshave developed. In other embodiments, treatment may be administered inthe absence of symptoms. For example, treatment may be administered to asusceptible individual prior to the onset of symptoms (e.g., in light ofa history of symptoms and/or in light of genetic or other susceptibilityfactors). Treatment may also be continued after symptoms have resolved,for example to prevent or delay their recurrence.

“Patient” or “subject”, as used herein, means an animal subject,preferably a mammalian subject, and particularly human subjects(including both male and female subjects, and including neonatal,infant, juvenile, adolescent, adult and geriatric subjects). Subjectsmay also include other mammalian subjects (e.g., dog, cat, horse, cow,sheep, goat, monkey, bird, etc.), for laboratory or veterinary purposes.

In some embodiments, treatment is provided to a subject havinghepatocellular carcinoma with altered FGFR4 and/or FGF19 (fibroblastgrowth factor 19) status.

In some embodiments, treatment may include or be performed inconjunction with analyzing FGFR4 and/or FGF19 status in a biologicalsample containing cells of said hepatocellular carcinoma, and if saidhepatocellular carcinoma exhibits an FGFR4 and/or FGF19 alteration,treating a subject with a treatment effective amount of an active agentas described herein.

“Altered status” as used herein with reference to FGFR4 and/or FGF19includes an increased expression thereof (e.g., increased levels of themRNA or increased levels of the protein), increased copy number in thegenome, and/or increased activity of the encoded protein as a result ofmutation, etc., as compared to a corresponding non-cancerous tissue. Insome embodiments, altered status of FGFR4 and/or FGF9 includes geneand/or encoded protein mutations that result in an increase in activityor are otherwise associated with a more aggressive form ofhepatocellular carcinoma.

“Expression” of FGFR4 and/or FGF19 means that a gene encoding the sameis transcribed, and preferably, translated. Typically, expression of acoding region will result in production of the encoded polypeptide.

The FGFR4 and FGF19 proteins are known, and their altered status and/orexpression may be measured using techniques standard in the art, e.g.,genomic analysis of mutations or copy number aberrations such as bynucleic acid amplification, sequencing analysis, and/orhybridization-based techniques, RNA expression analysis such as northernblot or qRT-PCR, western blot or other immunoblot or immunoassay,fluorescent activated cell sorting (FACS), etc.

In order that the invention described herein may be more fullyunderstood, the following examples are set forth. It should beunderstood that these examples are for illustrative purposes only andare not to be construed as limiting.

EXAMPLES

General:

Microwave heating was done using a Biotage Emrys Liberator or Initiatormicrowave. Column chromatography was carried out using an Isco Rf200d.Solvent removal was carried out using either a Büchi rotary evaporatoror a Genevac centrifugal evaporator. Preparative LC/MS was conductedusing a Waters autopurifier and 19×100 mm XTerra 5 micron MS C18 columnunder acidic mobile phase conditions. NMR spectra were recorded using aVarian 400 MHz spectrometer.

When the term “inerted” is used to describe a reactor (e.g., a reactionvessel, flask, glass reactor, and the like) it is meant that the air inthe reactor has been replaced with an essentially moisture-free or dry,inert gas (such as nitrogen, argon, and the like).

General methods and experimentals for preparing compounds of the presentinvention are set forth below. In certain cases, a particular compoundis described by way of example. However, it will be appreciated that ineach case a series of compounds of the present invention were preparedin accordance with the schemes and experimentals described below.

Preparative HPLC Conditions for the Purification of Target Compounds

Chromatography Conditions:

-   -   Instrument: Waters 2767-SQD Mass trigger Prep System    -   Column: Waters Xbridge C18 150 mm*19 mm*5 μm    -   Detector: VWD SQD    -   Flow Rate: 15 mL/min    -   Gradient Time:

Time(min) B % 0 5 7.5 70 8 95 11 95

Representative Mobile Phase:

-   -   1)    -   Mobile Phase: A: 0.1% TFA in water    -   Mobile Phase: B: ACN    -   2)    -   Mobile Phase: A: 0.1% NH₄HCO₃ in water    -   Mobile Phase: B: ACN    -   3)    -   Mobile Phase: A: 0.1% NH₄OAc in water    -   Mobile Phase: B: ACN    -   4)    -   Mobile Phase: A: 0.1% NH₄OH in water    -   Mobile Phase: B: ACN

Definitions

The following abbreviations have the indicated meanings:

ACN: Acetonitrile

Boc₂O: Di-tert-butyl dicarbonate

Brettphos:2-(Dicyclohexylphosphino)-3,6-dimethoxy-2′,4′,6′-triisopropyl-1,1′-biphenyl

tBuONa: Sodium tert-butoxide

CH₃I: Iodomethane

Cs₂CO₃: Cesium carbonate

DCC: N,N′-dicyclohexylcarbodiimide

DCM: Dichloromethane

DIEA: N,N-diisopropylethylamine

DIPEA: N,N-diisopropylethylamine

DMAP: 4-(Dimethylamino)pyridine

DME: Dimethyl ether

DMF: Dimethylformamide

DMSO: Dimethyl sulfoxide

EGTA: Ethylene glycol tetraacetic acid

ESI-MS: Electrospray ionization—mass spectrometry

EtOH: Ethanol

HATU: 1-[Bis(dimethylamino)methylene]-1H-1,2,3-trizolo[4,5-b]pyridinium3-oxid hexafluorophosphate

H₂SO₄: Sulfuric acid

iPrOH: Isopropanol

K₂CO₃: Potassium carbonate

KHMDS: Potassium bis(trimethylsilyl)amide

KOH: Potassium hydroxide

LCMS: Liquid chromatography—mass spectrometry

MeOH: Methanol

MsCl: Methansulfonyl chloride

NaBH₃CN: Sodium cyanoborohydride

NaBH(OAc)₃: Sodium triacetoxyborohydride

NH₄Cl: Ammonium chloride

NH₄HCO₃: Ammonium bicarbonate

NaI: Sodium iodide

NaNO₃: Sodium nitrate

NaOAc: Sodium acetate

nBuOH: n-Butanol

prep-HPLC: Preparative high-performance liquid chromatography

prep-TLC: Preparative thin layer chromatography

TBAF: Tetrabutylammonium fluoride

TBDMS-CL: tert-Butyldimethylsilyl chloride

TBSCl: tert-Butyldimethylsilyl chloride

TBSOTf: tert-Butyldimethylsilyl trifluoromethanesulfonate

TEA: Triethylamine

TESCl: Chlorotriethylsilane

TFA: Trifluoroacetic acid

THF: Tetrahydrofuran

Ti(O^(i)Pr)₄: Titanium isopropoxide

TLC: Thin-layer chromatography

PPTS: Pyridinium p-toluenesulfonate

PE: Petroleum ether

PEG: Poly(ethylene glycol)

PtO₂: platinum dioxide

EtOAc: Ethyl acetate

Pd/C: Palladium (0) on carbon

Pd₂(dba)₃: Tris(dibenzylideneacetone) dipalladium(0)

Pd(dppf)₂Cl₂:[1,1′-Bis(diphenylphosphino)ferrocene]dichloropalladium(II)

Ruphos: 2-Dicyclohexylphosphino-2′,6′-diisopropoxybiphenyl

Xantphos: 4,5-Bis(diphenylphosphino)-9,9-dimethylxanthene

Materials:

The following compounds are commercially available and/or can beprepared in a number of ways well known to one skilled in the art oforganic synthesis. More specifically, disclosed compounds can beprepared using the reactions and techniques described herein. In thedescription of the synthetic methods described below, it is to beunderstood that all proposed reaction conditions, including choice ofsolvent, reaction atmosphere, reaction temperature, duration of theexperiment, and workup procedures, can be chosen to be the conditionsstandard for that reaction, unless otherwise indicated. It is understoodby one skilled in the art of organic synthesis that the functionalitypresent on various portions of the molecule should be compatible withthe reagents and reactions proposed. Substituents not compatible withthe reaction conditions will be apparent to one skilled in the art, andalternate methods are therefore indicated. The starting materials forthe examples are either commercially available or are readily preparedby standard methods from known materials.

SYNTHESIS AND TESTING OF EXAMPLE COMPOUNDS

Compounds of Table 1 were prepared by the Procedures of 2A-2L.

TABLE 1 Compound FGFR4 FGFR1 Structure # IC₅₀ (μM) IC₅₀ (μM)

100 <0.001 >10.0

102 0.853 >10.0

103 0.673 >10.0

105 0.201 >10.0

107 <0.001 0.720

108 <0.001 0.173

110 0.027 >50.0

111 <0.001 1.280

112 <0.001 2.600

113 <0.001 0.621

114 <0.001 0.042

116 0.004 >20.0

120 <0.001 1.190

121 <0.001 1.020

122 <0.001 0.324

123 <0.001 3.890

124 <0.001 0.164

125 <0.001 2.860

126 <0.001 0.601

127 <0.001 3.200

128 0.033 1.680

129 0.063 >10.0

130 0.002 >10.0

131 <0.001 1.170

132 <0.001 0.495

133 0.004 >10.0

135 <0.001 2.027

136 <0.001 0.112

137 <0.001 >10.0

139 0.006 >20.0

140 <0.001 >20.0

141 0.002 >20.0

142 <0.001 >10.0

143 0.010 0.155

144 <0.001 0.105

145 <0.001 2.190

147 <0.001 5.940

148 <0.001 6.440

149 <0.001 7.700

150 <0.001 2.290

151 <0.001 0.197

152 0.014 0.297

154 <0.001 0.777

155 <0.001 8.030

156 <0.001 1.950

157 <0.001 0.131

158 <0.001 2.152

159 <0.001 0.115

160 <0.001 0.660

161 <0.001 2.694

162 <0.001 >10.0

163 <0.001 0.519

164 <0.001 3.510

165 <0.001 7.370

166 <0.001 4.920

167 <0.001 >10.0

168 <0.001 1.030

170 0.008 >10.0

171 0.003 3.920

172 0.011 7.370

175 0.015 >10.0

181 <0.001 0.224

183 0.180 6.210

184 0.089 6.475

185 0.013 >10.0

186 0.345 >10.0

Procedure 2A: Example—100

N-(2-{6-[3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-methyl-ureido]-pyrimidin-4-ylamino}-phenyl)-acrylamide

a. N-(3,5-Dimethoxy-phenyl)-acetamide

To a solution of 3,5-dimethoxy-phenylamine (20 g, 0.131 mol) in toluene(110 mL) was added acetic anhydride (14 g, 0.137 mmol) at roomtemperature. The resulting mixture was stirred for 18 hours at roomtemperature. PE (55 mL) was added, the precipitate was filtered andwashed with PE (100 mL) to obtain the title compound (24.2 g, yield:95%). ¹H-NMR (400 MHz, CDCl₃) δ 2.16 (s, 3H), 3.77 (s, 6H), 6.23 (s,1H), 6.75 (s, 2H), 7.20 (s, 1H).

b. N-(2,6-Dichloro-3,5-dimethoxy-phenyl)-acetamide

To a solution of N-(3,5-dimethoxy-phenyl)-acetamide (5 g, 25.6 mmol) inACN (75 mL) was added sulfuryl chloride (6.9 g, 51.2 mmol) at 0° C.under nitrogen atmosphere. The resulting mixture was stirred for 30minutes at this temperature and quenched with saturated aqueous NaHCO₃(40 mL). The precipitate was filtered, washed with water and dried toobtain the title compound (2.3 g, yield: 34%). ¹H-NMR (400 MHz, CDCl₃) δ2.25 (s, 3H), 3.86 (s, 6H), 6.54 (s, 1H), 6.90 (s, 1H).

c. 2,6-Dichloro-3,5-dimethoxy-phenylamine

A solution of N-(2,6-dichloro-3,5-dimethoxy-phenyl)-acetamide (3.6 g,13.7 mmol) in EtOH (130 mL) and KOH (2M, 75 mL) was heated to reflux for24 hours. The reaction was cooled to 0° C. and stirred for 1 hour atthis temperature. The precipitate was filtered and dried to obtain thetitle compound (2.3 g, yield: 76%). ¹H-NMR (400 MHz, CDCl₃) δ 3.90 (s,6H), 4.57 (bs, 2H), 6.05 (s, 1H).

d. 2,4-Dichloro-3-isocyanato-1,5-dimethoxy-benzene

A mixture of 2,6-dichloro-3,5-dimethoxy-phenylamine (500 mg, 2.25 mmol),triphosgene (335 mg, 1.12 mmol) and TEA (342 mg, 3.38 mmol) in dioxane(15 mL) was heated to 130° C. for 2 hours under microwave. The reactionwas concentrated and the residue was purified by flash chromatography onsilica eluting with DCM to obtain the title compound (450 mg, yield:80%). ¹H-NMR (400 MHz, CDCl₃) δ 3.92 (s, 6H), 6.42 (s, 1H).

e. (6-Chloro-pyrimidin-4-yl)-methyl-amine

To a solution of 4,6-dichloro-pyrimidine (7.45 g, 50 mmol) in iPrOH (50mL) was added a solution of methyl amine in THF (2M, 30 mL, 60 mmol) atroom temperature. The resulting mixture was stirred for 18 hours. Themixture was concentrated and the residue was purified by flashchromatography on silica eluting with DCM:EtOAc=6:1-1:1 to obtain thetitle compound (4.4 g, yield: 62%) as a white solid. ¹H NMR (400 MHz,CDCl₃) δ 2.96 (d, 3H), 5.22-5.36 (bs, 1H), 6.35 (s, 1H), 8.35 (s, 1H);MS (ESI): 144 [M+H]⁺.

f. N-Methyl-N′-(2-nitro-phenyl)-pyrimidine-4,6-diamine

A mixture of (6-chloro-pyrimidin-4-yl)-methyl-amine (1 g, 7 mmol),2-nitro-phenylamine (965 mg, 7 mmol), Brettphos (279 mg, 0.35 mmol) andtBuONa (2 g, 21 mmol) in DME (50 mL) was heated to 90° C. for 1 hourunder nitrogen atmosphere. The reaction was concentrated, and theresidue was purified by flash chromatography on silica eluting withDCM:EtOAc=10:1-1:1 to obtain the title compound (600 mg, yield: 35%) asa yellow solid. ¹H NMR (400 MHz, CDCl₃) δ 2.94 (d, 3H), 4.99 (bs, 1H),5.82 (s, 1H), 7.04 (t, 1H), 7.60 (t, 1H), 8.21 (d, 1H), 8.33 (s, 1H),8.75 (d, 1H), 9.91 (s, 1H); MS (ESI): 246 [M+H]⁺.

g.3-(2,6-Dichloro-3,5-dimethoxy-phenyl)-1-methyl-1-[6-(2-nitro-phenylamino)-pyrimidin4-yl]-urea

To a solution of N-methyl-N′-(2-nitro-phenyl)-pyrimidine-4,6-diamine(150 mg, 0.61 mmol) in THF (15 mL) was added NaH (60%, 60 mg, 1.5 mmol)at 0° C., and the mixture was stirred for 30 minutes at roomtemperature. A solution of2,4-dichloro-3-isocyanato-1,5-dimethoxy-benzene (180 mg, 0.73 mmol) wasadded dropwise at room temperature. The resulting mixture was stirredfor 2 hours. Water (2 mL) was added to quench the reaction. The mixturewas concentrated, and the residue was purified by flash chromatographyon silica eluting with DCM:EtOAc=6:1-1:1 to obtain the title compound(54 mg, yield: 18%) as a yellow solid. ¹H NMR (400 MHz, DMSO-d6) δ 3.38(s, 3H), 3.93 (s, 6H), 6.75 (s, 1H), 6.91 (s, 1H), 7.34 (t, 1H), 7.72(t, 1H), 7.79 (d, 1H), 8.01 (d, 1H), 8.38 (s, 1H), 9.99 (s, 1H), 11.78(s, 1H); MS (ESI): 493 [M+H]⁺.

h.1-[6-(2-Amino-phenylamino)-pyrimidin-4-yl]-3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-methyl-urea

To a solution of3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-methyl-1-[6-(2-nitro-phenylamino)-pyrimidin-4-yl]-urea(50 mg, 0.1 mmol) in THF (10 mL) and MeOH (10 mL) was added Raney-Ni(suspension in water) at room temperature, the resulting mixture wasstirred for 2 hours under hydrogen atmosphere. The reaction was filteredand concentrated to obtain the title compound (38 mg, yield: 82%), whichwas used directly in the next step. ¹H NMR (400 MHz, CDCl₃) δ 3.28 (s,3H), 3.85 (s, 2H), 3.94 (s, 6H), 5.86 (s, 1H), 6.52 (s, 1H), 6.78-6.87(m, 3H), 7.16-7.20 (m, 2H), 8.39 (s, 1H), 12.62 (s, 1H); MS (ESI): 463[M+H]⁺.

i.N-(2-{6-[3-(2,6-Dichloro-3,5-dimethoxy-phenyl)-1-methyl-ureido]-pyrimidin-4-ylamino}-phenyl)-acrylamide

To a solution of1-[6-(2-amino-phenylamino)-pyrimidin-4-yl]-3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-methyl-urea(25 mg, 0.05 mmol) in THF (10 mL) was added a solution of acryloylchloride in THF (20 mg/mL, 0.5 mL, 0.1 mmol) at −10° C., and theresulting mixture was stirred for 1 hour at this temperature. MeOH (1mL) was added to quench the reaction. The mixture was concentrated andthe residue was purified by prep-TLC to obtain the title compound (12mg, yield: 43%). ¹H NMR (400 MHz, DMSO-d6) δ 3.26 (s, 3H), 3.94 (s, 6H),5.74 (d, 1H), 6.24 (d, 1H), 6.37 (s, 1H), 6.47-6.54 (m, 1H), 6.90 (s,2H), 7.20 (d, 2H), 7.56-7.58 (m, 1H), 7.66-7.68 (m, 1H), 8.38 (s, 1H),9.99 (s, 1H), 9.70 (s, 1H), 11.99 (s, 1H); MS (ESI): 517 [M+H]⁺.

Compounds 102, 103 and 105 were synthesized in a similar manner ascompound 100.

Procedure 2B: Example—107

N-(2-{6-[3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-pyridin-3-ylmethyl-ureido]-pyrimidin-4-ylamino}-phenyl)-acrylamide

a. (6-Chloro-pyrimidin-4-yl)-pyridin-3-ylmethyl-amine

To a solution of 4,6-dichloro-pyrimidine (1 g, 6.71 mmol) in dioxane (20mL) was added a solution of pyridin-3-yl-methylamine (745 mg, 6.9 mmol)at room temperature. The resulting mixture was stirred at roomtemperature overnight. The mixture was concentrated and the residue waspurified by flash chromatography on silica to obtain the title compound(680 mg, yield: 46%). MS (ESI): 221 [M+H]⁺.

b. N-(2-Nitro-phenyl)-N′-pyridin-3-ylmethyl-pyrimidine-4,6 diamine

A degassed mixture of (6-chloro-pyrimidin-4-yl)-pyridin-3-ylmethyl-amine(300 mg, 1.36 mmol), 2-nitro-phenylamine (188 mg, 1.36 mmol), Pd₂(dba)₃(128 mg, 0.14 mmol), Xantphos (161 mg, 0.28 mmol) and Cs₂CO₃ (913 mg,2.8 mmol) in toluene (10 mL) was heated at 100° C. for 4 hours. Thereaction was concentrated, and the residue was purified by flashchromatography on silica to obtain the title compound (150 mg, yield:34%). MS (ESI): 323 [M+H]⁺.

c.3-(2,6-Dichloro-3,5-dimethoxy-phenyl)-1-[6-(2-nitro-phenylamino)-pyrimidin-4-yl]-1-pyridin-3-ylmethyl-urea

To a solution ofN-(2-nitro-phenyl)-N′-pyridin-3-ylmethyl-pyrimidine-4,6-diamine (150 mg,0.467 mmol) in THF (15 mL) was added NaH (60%, 48 mg, 1.2 mmol) at 0°C., and the mixture was stirred for 30 minutes at room temperature. Asolution of 2,4-dichloro-3-isocyanato-1,5-dimethoxy-benzene (procedure2A, steps a-d; 180 mg, 0.73 mmol) was added dropwise at roomtemperature. The resulting mixture was stirred for 2 hours. Water (2 mL)was added to quench the reaction. The mixture was concentrated, and theresidue was purified by flash chromatography on silica to obtain thetitle compound (85 mg, yield: 32%). MS (ESI): 570 [M+H]⁺.

d.1-[6-(2-Amino-phenylamino)-pyrimidin-4-yl]-3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-pyridin-3-ylmethyl-urea

A mixture of3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-[6-(2-nitro-phenylamino)-pyrimidin-4-yl]-1-pyridin-3-ylmethyl-urea(85 mg, 0.149 mmol) and Fe (84 mg, 1.5 mmol) in AcOH (5 mL) was heatedat 50° C. for 2 hours. The reaction mixture was filtered and thefiltrate was concentrated in vacuo to give crude product, which waspurified by silica gel column chromatography to afford the titlecompound (53 mg, yield: 66%). MS (ESI): 540 [M+H]⁺.

e. N-(2-{6-[3-(2,6-Dichloro-3,5-dimethoxy-phenyl)-1-pyridin-3-ylmethyl-ureido]-pyrimidin-4-ylamino}-phenyl-acrylamide

To a solution of1-[6-(2-amino-phenylamino)-pyrimidin-4-yl]-3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-pyridin-3-ylmethyl-urea(53 mg, 0.1 mmol) in THF (10 mL) was added a solution of acryloylchloride in THF (20 mg/mL, 0.5 mL, 0.1 mmol) at −10° C., and the mixturewas stirred for 1 hour at this temperature. MeOH (1 mL) was added toquench the reaction. The mixture was concentrated and the residue waspurified by prep-TLC to obtain the title compound (9 mg, yield: 15%). ¹HNMR (400 MHz, CDCl₃) δ 3.84 (s, 6H), 5.01 (s, 2H), 5.69 (d, 1H), 5.75(s, 1H), 6.10 (dd, 1H), 6.34 (d, 1H), 6.47 (s, 1H), 7.00 (d, 1H),7.09-7.24 (m, 2H), 7.28 (t, 1H), 7.32 (s, 1H), 7.47 (d, 1H), 7.69-7.71(m, 2H), 8.31-8.34 (m, 2H), 8.40-8.42 (m, 1H), 12.60 (s, 1H); MS (ESI):594 [M+H]⁺.

Procedure 2C: Example—108

N-(2-(6-(3-(2,6-Dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-ylamino)-5-(4-ethylpiperazin-1-yl)-phenyl)acrylamide

a. tert-Butyl 4-bromo-2-nitrophenylcarbamate

A mixture of 4-bromo-2-nitroaniline (4 g, 18.4 mmol), (Boc)₂O (4.4 g,20.24 mmol) in THF (50 mL) was heated under reflux overnight. Themixture was concentrated and the residue was purified by flashchromatography on silica eluting with PE:EtOAc=20:1 to obtain the titlecompound (5.4 g, yield: 93%). MS (ESI): 317, 319 [M+H]⁺.

b. tert-Butyl 4-(4-ethylpiperazin-1-yl)-2-nitrophenylcarbamate

A degassed mixture of tert-butyl 4-bromo-2-nitrophenylcarbamate (5.4 g,17 mmol), 1-ethylpiperazine (2.91 g, 25.5 mmol), Pd₂(dba)₃ (2.1 g, 3.4mmol), xantphos (3.92 g, 6.8 mmol) and Cs₂CO₃ (11.1 g, 34 mmol) intoluene (85 mL) was heated at 100° C. for 4 hours. The reaction wasconcentrated, and the residue was purified by flash chromatography onsilica eluting with MeOH:DCM=1:50-1:20 to obtain the title compound (3.3g, yield: 55%). MS (ESI): 351 [M+H]⁺.

c. 4-(4-Ethylpiperazin-1-yl)-2-nitroaniline

To a solution of tert-butyl4-(4-ethylpiperazin-1-yl)-2-nitrophenylcarbamate (3.3 g, 9.43 mmol) inDCM (50 mL) was added TFA (20 mL) at 0° C., the resulting mixture wasstirred for 3 hours at rt. After removal of all volatiles in vacuo, theresidue was re-dissolved in DCM, neutralized with saturated aqueousK₂CO₃ and extracted with DCM. The combined extracts were concentrated toobtain the title compound (2.1 g, yield: 90%), which was used directlyin the next step. ¹H NMR (400 MHz, DMSO-d6) δ 1.02 (t, 3H), 2.36 (q,2H), 2.47-2.49 (m, 4H) 2.97-3.00 (m, 4H), 6.97 (d, 1H), 7.20 (s, 2H),7.25 (s, 1H), 7.34 (dd, 1H); MS (ESI): 251 [M+H]⁺.

d.N-(4-(4-ethylpiperazin-1-yl)-2-nitrophenyl)-N⁶-methylpyrimidine-4,6-diamine

A degassed mixture of 4-(4-ethylpiperazin-1-yl)-2-nitroaniline (2.1 g,8.4 mmol), 6-chloro-N-methylpyrimidin-4-amine (Procedure 2A, step e; 1.2g, 8.4 mmol), Pd₂(dba)₃ (1.54 g, 1.68 mmol), xantphos (1.94 g, 3.36mmol) and Cs₂CO₃ (5.48 g, 16.8 mmol) in toluene (45 mL) was heated at100° C. for 1 hour. The reaction was concentrated, and the residue waspurified by flash chromatography on silica eluting withMeOH:DCM=1:40-1:20 to obtain the title compound (870 mg, yield: 29%). MS(ESI): 358 [M+H]⁺.

e.3-(2,6-Dichloro-3,5-dimethoxyphenyl)-1-(6-(4-(4-ethylpiperazin-yl)-2-nitrophenylamino)pyrimidin-4-yl)-1-methylurea

To a solution ofN4-(4-(4-ethylpiperazin-1-yl)-2-nitrophenyl)-N6-methylpyrimidine-4,6-diamine(870 mg, 2.44 mmol) in THF (15 mL) was added NaH (60%, 200 mg, 5 mmol)at 0° C., and the mixture was stirred for 30 minutes at roomtemperature. A solution of2,4-dichloro-3-isocyanato-1,5-dimethoxy-benzene (Procedure 2A, stepsa-d; 908 mg, 3.66 mmol) in THF was added dropwise at 0° C. The resultingmixture was stirred at room temperature for 2 hours. Saturated aqueousNH₄Cl solution (2 mL) was added to quench the reaction. The mixture wasconcentrated and extracted with DCM. The combined extracts were washedwith brine, dried over anhydrous Na₂SO₄, and concentrated to give thecrude product, which was purified by flash chromatography on silica toobtain the title compound (330 mg, yield: 21%) as a red oil. ¹H NMR (400MHz, CDCl₃) δ 1.44 (t, 3H), 3.01 (t, 2H), 3.21 (q, 2H), 3.41-3.49 (m,5H), 3.73-3.80 (m, 4H), 3.92 (s, 6H), 6.27 (s, 1H), 6.55 (s, 1H), 7.25(d, 1H), 7.69 (s, 1H), 8.32 (d, 1H), 8.52 (s, 1H), 10.28 (br s, 1H),12.05 (br s, 1H); MS (ESI): 605 [M+H]⁺.

f.1-(6-(2-Amino-4-(4-ethylpiperazin-1-yl)phenylamino)pyrimidin-4-yl)-3-(2,6-dichloro-3,5-dimethoxyphenyl-1-methylurea

To a solution of3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-(6-(4-(4-ethylpiperazin-1-yl)-2-nitrophenylamino)pyrimidin-4-yl)-1-methylurea(330 mg, 0.546 mmol) in THF (20 mL) and MeOH (20 mL) was added Raney-Ni(suspension in water) at room temperature, the resulting mixture wasstirred for 3 hours under hydrogen atmosphere (1 atm). The reaction wasfiltered and concentrated. The residue was washed twice with MeOH toobtain the title compound (280 mg, purity: 90%), which was used directlyin the next step. MS (ESI): 575 [M+H]⁺

g.N-(2-(6-(3-(2,6-Dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-ylamino)-5-(4-ethylpiperazin-1-yl)phenyl)acrylamide

To a solution of1-(6-(2-amino-4-(4-ethylpiperazin-1-yl)phenylamino)pyrimidin-4-yl)-3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylurea(280 mg, purity: 90%, 0.44 mmol) in THE (30 mL) was added a solution ofacryloyl chloride in THF (20 mg/mL, 2 mL, 0.44 mmol) at −10° C., and theresulting mixture was stirred for 1 hour at this temperature. MeOH (1mL) was added to quench the reaction. The mixture was concentrated andthe residue was purified by prep-HPLC and prep-TLC to obtain the titlecompound (20 mg, yield: 7%). ¹H NMR (400 MHz, CDCl₃) δ 1.31 (t, 3H),2.65 (q, 2H), 2.62-2.68 (m, 4H), 3.27 (s, 3H), 3.36-3.38 (m, 4H), 3.91(s, 6H), 5.76 (d, 1H), 5.90 (s, 1H), 6.24 (dd, 1H), 6.41 (d, 1H), 6.52(s, 1H), 6.74 (dd, 1H), 7.07 (br s, 1H), 7.23 (d, 1H), 7.72 (br s, 1H),7.98 (br s, 1H), 8.37 (s, 1H), 12.52 (s, 1H); MS (ESI): 629 [M+H]⁺.

Example—110

N-(2-(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-ylamino)-phenyl)-2-fluoroacrylamide

The compound was synthesized following the approach outlined inProcedure 2A (Example 100), modifying step (i) to the followingprocedure: To a solution of1-[6-(2-amino-phenylamino)-pyrimidin-4-yl]-3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-methyl-urea(130 mg, mixed with tetrachloro aniline) and DCC (118 mg, 0.56 mmol) inchloroform (100 mL) was added a solution of 2-fluoroacrylic acid (50 mg,0.56 mmol) in chloroform (50 mL) at 0° C., and the resulting mixture wasstirred at room temperature overnight. Water (I mL) was added to quenchthe reaction. The mixture was concentrated and the residue was purifiedby reverse phase column and prep-TLC to obtain the title compound (4 mg,yield: 5%). ¹H NMR (400 MHz, CDCl₃) δ 12.17 (s, 1H), 8.45 (s, 1H), 8.35(s, 1H), 7.84 (d, 1H), 7.37 (d, 1H), 7.29 (t, 1H), 7.26 (t, 1H), 6.47(s, 1H), 5.94 (s, 1H), 5.78 (dd, 1H), 5.21 (dd, 1H), 3.85 (s, 6H), 3.25(s, 3H); MS (ESI): 535 [M+H]⁺.

Procedure 2E: Example—111

N-(2-(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-ylamino)-5-(1-ethylpiperidin-4-yl)phenyl)acrylamide

a. tert-butyl 4-(4-amino-3-nitrophenyl)-5,6-dihydropyridine-1(2H)-carboxylate

To a degassed mixture of 4-bromo-2-nitroaniline (1 g, 4.6 mmol),tert-butyl4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5,6-dihydropyridine-1(2H)-carboxylate(1.42 g, 4.6 mmol), tripotassium phosphate trihydrate (3.9 g, 14.64mmol) in dioxane and water (30 mL, 8:1) was added Pd(dppf)₂Cl₂ (337 mg,0.46 mmol). The mixture was refluxed at 110° C. for 3 hours.

Filtration and concentration gave crude product, which was purified bysilica gel column chromatography to afford the title compound (1.1 g,yield: 75%). MS (ESI): 320 [M+H]⁺.

b.1-(6-chloropyrimidin-4-yl)-3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-((2-(trimethylsilyl)ethoxy)methyl) urea

To a solution of 6-chloro-N-methylpyrimidin-4-amine (Procedure 2A, stepe; 460 mg, 3.21 mmol) in DMF (15 mL) was added NaH (60%, 193 mg, 4.81mmol) at 0° C., and the mixture was stirred for 30 minutes at roomtemperature. A solution of2,4-dichloro-3-isocyanato-1,5-dimethoxy-benzene (Procedure H, steps a-d;1.03 g, 4.17 mmol) in DMF (5 mL) was added dropwise at room temperature.The resulting mixture was stirred for 0.5 hour. SEMCl (804 mg, 4.81mmol) in DMF (2 mL) was added. The reaction mixture was stirred at roomtemperature for 1 hour. Saturated aqueous NH₄Cl was added to quench thereaction. The mixture was diluted with water and extracted with EtOAc.The combined extracts were washed with water and brine, dried overanhydrous Na₂SO₄ and filtered. The filtrate was evaporated under vacuumto give crude product, which was purified by flash chromatography onsilica to obtain the title compound (470 mg, yield: 28%). MS (ESI): 521[M+H]⁺.

c.tert-butyl-(4-(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-((2-(trimethylsilyl)ethoxy)methyl)ureido)pyrimidin-4-ylamino)-3-nitrophenyl)-5,6-dihydropyridine-1(2H)-carboxylate

A degassed mixture of1-(6-chloropyrimidin-4-yl)-3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-((2-(trimethylsilyl)ethoxy)methyl)urea(470 mg, 0.9 mmol), tert-butyl4-(4-amino-3-nitrophenyl)-5,6-dihydropyridine-1(2H)-carboxylate (320 mg,1 mmol), Pd₂(dba)₃ (92 mg, 0.1 mmol), xantphos (115 mg, 0.2 mmol) andCs₂CO₃ (652 mg, 2 mmol) in toluene (10 mL) was heated at 100° C. for 5hours. The reaction was concentrated, and the residue was purified byflash chromatography on silica to obtain the title compound (400 mg,yield: 57%). MS (ESI): 804 [M+H]⁺.

d.tert-butyl-4-(3-amino-4-(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-((2-(trimethylsilylethoxy)methy)ureido)pyrimidin-4-ylamino)phenyl)piperidine-1-carboxylate

To a solution oftert-butyl-4-(4-(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-((2-(trimethylsilyl)ethoxy)methyl)ureido)pyrimidin-4-ylamino)-3-nitrophenyl)-5,6-dihydropyridine-1(2H)-carboxylate(380 mg, 0.473 mmol) in MeOH (10 mL) was added PtO₂ (38 mg, 10% wt) andone drop of chlorobenzene at room temperature, the resulting mixture wasstirred under hydrogen atmosphere (1 atm) overnight. The reaction wasfiltered and concentrated. The residue was purified by flashchromatography on silica to obtain the title compound (130 mg, yield:37%). MS (ESI): 776 [M+H]⁺.

e.N-(2-(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-ylamino)-5-(piperidin-4-yl)phenyl)acrylamideTFA salt

To a solution oftert-butyl-4-(3-amino-4-(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-((2-(trimethylsilyl)ethoxy)methyl)ureido)pyrimidin-4-ylamino)phenyl)piperidine-1-carboxylate(130 mg, 0.168 mmol) in THF (15 mL) was added a solution of acryloylchloride (10 mg/mL, 1.7 mL, 0.19 mmol) dropwise at −10° C., and theresulting mixture was stirred at 0° C. for 1 hour. LC-MS showed that thereaction was complete. MeOH (5 mL) was added to quench the reaction, andthe reaction was concentrated. The residue in DCM (2 mL) was addeddropwise to a mixture of DCM/TFA (2/1, v/v, 3 mL). The mixture wasstirred at room temperature for 1 hour and then concentrated undervacuum. The residue (50 mg, quant.) was used directly for the next stepwithout further purification. MS (ESI): 600 [M+H]⁺.

f.N-(2-(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-ylamino)-5-(1-ethylpiperidin-4-yl)phenyl)acrylamide

To a solution ofN-(2-(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-ylamino)-5-(piperidin-4-yl)phenyl)acrylamideTFA salt (35 mg, 0.049 mmol) in EtOH (1 mL), was added NaOAc (4 mg, 0.05mmol) and aqueous acetaldehyde (1 mL, 0.9 mmol, 40%). After the mixturewas stirred at room temperature for 1 hour, NaBH₃CN (12 mg, 0.18 mmol)was added, and the solution was stirred at room temperature for another3 hours. After removal of all volatiles in vacuo, the residue waspartitioned between DCM and water. The aqueous layer was extracted withchloroform twice. The combined extracts were washed with brine, driedover anhydrous Na₂SO₄ and filtered. The filtrate was evaporated undervacuum to give crude product, which was purified by prep-HPLC to affordthe title compound (3 mg, yield: 10%). ¹H NMR (400 MHz, MeOH-d4) δ 8.38(s, 1H), 7.68 (s, 1H), 7.54 (d, 1H), 7.24 (dd, 1H), 6.83 (s, 1H),6.46-6.35 (m, 3H), 5.81 (d, 1H), 3.97 (s, 6H), 3.74-3.70 (m, 2H), 3.37(s, 3H), 3.26 (q, 2H), 3.17-3.11 (m, 2H), 2.99-2.96 (m, 1H), 2.26-2.22(m, 2H), 2.06-1.99 (m, 2H), 1.43 (t, 3H); MS (ESI): 628 [M+H]⁺.

Example—112

N-(2-(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-ylamino)-5-(1-methylpiperidin-4-yl)phenyl)acrylamide

The compound was synthesized following the approach outlined inProcedure 2E (Example 111), substituting formaldehyde in step (f) toafford the title compound (1.5 mg, yield: 11.6%). ¹H NMR (400 MHz,MeOH-d4) δ 8.26 (s, 1H), 7.55 (s, 1H), 7.41 (d, 1H), 7.11 (d, 1H), 6.71(s, 1H), 6.34-6.22 (m, 3H), 5.68 (d, 1H), 3.84 (s, 6H), 3.55-3.52 (m,2H), 3.25 (s, 3H), 3.21-3.08 (m, 2H), 2.90-2.83 (m, 4H), 2.11-2.08 (m,2H), 1.89-1.85 (m, 2H); MS (ESI): 614 [M+H]⁺.

Procedure 2F: Example—113

N-(2-{6-[3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-methyl-ureido]-pyrimidin-4-ylamino}-5-dimethylaminomethyl-phenyl)-acrylamide

a. 4-Fluoro-3-nitro-benzaldehyde

To a stirred solution of (4-fluoro-3-nitro-phenyl)-methanol (750 mg, 4.4mmol) in DCM (40 mL) at 0° C. was added Dess-Martin Reagent (3.0 g, 7mmol). The solution was stirred at room temperature for 4 hours. TLCshowed disappearance of starting material. The reaction was quenchedwith 10% NaHCO₃ and 10% Na₂S₂O₃ aqueous solution and DCM layer separatedand washed with water (100 mL) and brine (50 mL). The reaction wasconcentrated, and the residue was purified by flash chromatography onsilica to obtain the title compound (570 mg, yield: 75%). ¹H-NMR (400MHz, DMSO-d6) δ 10.09 (d, 1H), 8.36 (t, 1H), 8.06 (dd, 1H), 7.97 (m,1H).

b. 4-Amino-3-nitrobenzaldehyde

To a solution of 4-fluoro-3-nitro-benzaldehyde (570 mg, 3.3 mmol) in THF(20 mL) was added NH₄OH (5 mL). The reaction mixture was stirred at roomtemperature for 1 hour. The resulting yellow solid was collected andwashed with water, dried under vacuum to give the title compound (300mg, yield: 53%). ¹H-NMR (300 MHz, DMSO-d6) δ 9.76 (s, 1H), 8.57 (d, 1H),8.18 (br s, 2H), 7.80 (dd, 1H), 7.10 (d, 1H).

c. 4-Dimethylaminomethyl-2-nitro-phenylamine

To a stirred solution of dimethylamine (4.0 mL, 2 M, 8.0 mmol) in MeOH(4 mL) was added Ti(O^(i)Pr)₄ (1.15 g, 4 mmol) and the solution wasstirred at room temperature for 15 minutes Then4-amino-3-nitro-benzaldehyde (160 mg, 1.0 mmol) in MeOH (2 mL) was addedand the solution was stirred at room temperature overnight. Then NaBH₄(78 mg, 2 mmol) was added and the solution was stirred at roomtemperature for 1 hour. LCMS showed major product peak. The solution wasdiluted with EtOAc (60 mL) and washed with water (2×100 mL) and brine(50 mL), dried over anhydrous Na₂SO₄. The solution was evaporated todryness and 130 mg of crude product was collected, which was used forthe next step without further purification. III NMR (400 MHz, DMSO-d6) δ7.82 (s, 1H), 7.35 (br s, 2H), 7.31 (dd, 1H), 6.97 (d, 1H), 3.26 (s,2H), 2.12 (s, 6H).

d.1-(2,6-Dichloro-3,5-dimethoxy-phenyl)-3-[6-(4-dimethylaminomethyl-2-nitro-phenylamino)-pyrimidin-4-yl]-3-methyl-1-(2-trimethylsilanyl-ethoxymethyl)-urea

To a stirred solution of1-(6-chloro-pyrimidin-4-yl)-3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-methyl-3-(2-trimethylsilanyl-ethoxymethyl)-urea(procedure L, step b; 260 mg, 0.5 mmol) in toluene (5 mL) was added4-dimethylaminomethyl-2-nitro-phenylamine (100 mg, 0.5 mmol), Cs₂CO₃(400 mg, 1.25 mmol), Pd₂(dba)₃ (46 mg, 0.05 mmol), xantphos (90 mg, 0.15mmol). The solution was stirred at 100° C. overnight. LCMS showed majorproduct peak. The solution was evaporated with silica gel and purifiedby flash chromatography on silica eluting with EtOAc (w(/0.5% TEA):MeOH(w/0.5% TEA)=10-10:0.5 to afford the desired product (100 mg, yield:30%). MS (ESI): 680 [M+H]⁺.

e.[6-(2-Amino-4-dimethylaminomethyl-phenylamino)-pyrimidin-4-yl]-3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-methyl-3-(2-trimethylsilanyl-ethoxymethyl)-urea

To a stirred solution of1-(2,6-dichloro-3,5-dimethoxy-phenyl)-3-[6-(4-dimethylaminomethyl-2-nitro-phenylamino)-pyrimidin-4-yl]-3-methyl-1-(2-trimethylsilanyl-ethoxymethyl)-urea(100 mg, 0.15 mmol) in MeOH (10 mL) was added 4 drops of chlorobenzeneand then PtO₂ (30 mg, 30% wt). The solution was stirred under hydrogenatmosphere at room temperature overnight. The reaction was filtered andconcentrated. The residue was taken to the next step without furtherpurification. MS (ESI): 650 [M+H]⁺.

f.1-[6-(2-Amino-4-dimethylaminomethyl-phenylamino)-pyrimidin-4-yl]-3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-methyl-urea

To a stirred solution of1-[6-(2-amino-4-dimethylaminomethyl-phenylamino)-pyrimidin-4-yl]-3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-methyl-3-(2-trimethylsilanyl-ethoxymethyl)-ureain anhydrous DCM (10 mL) was added TFA (10 mL) at room temperature. Thesolution was stirred at room temperature for 3 hours. LCMS showed majorproduct peak. The solution was evaporated to dryness, diluted with DCM(40 mL) and washed with 10% saturated Na₂CO₃ (10 mL). The DCM layer wasdried over anhydrous Na₂SO₄. Concentration under vacuum gave crudeproduct, which was purified by silica gel column chromatography (10%MeOH/DCM with 0.5% Et₃N) to afford the title compound (45 mg, yield: 58%in two steps). MS (ESI): 520 [M+H]⁺,

g.N-(2-{6-[3-(2,6-Dichloro-3,5-dimethoxy-phenyl)-1-methyl-ureido]-pyrimidin-4-ylamino}-5-dimethylaminomethyl-phenyl)-acrylamide

To a stirred solution of1-[6-(2-amino-4-dimethylaminomethyl-phenylamino)-pyrimidin-4-yl]-3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-methyl-urea(45 mg, 0.11 mmol) in THF (40 mL) at −10° C. was added acryloyl chloride(30 mg, 0.33 mmol) in THF (3 mL). The solution was stirred at −10° C.for 5 hours. LCMS showed major product peak. The reaction was quenchedwith MeOH (3 mL) and evaporated. The residue was purified by prep-HPLC(water/ACN in NH₄HCO₃ condition) to afford the title compound (6 mg,yield: 15%). ¹H-NMR (400 MHz, MeOH-d₄) δ 8.25 (s, 1H), 7.47 (d, 1H),7.44 (s, 1H), 7.16 (dd, 1H), 6.70 (s, 1H), 6.33-6.20 (m, 3H), 5.67 (dd,1H), 3.84 (s, 6H), 3.43 (s, 2H) 3.22 (s, 3H), 2.20 (s, 6H); MS (ESI):574 [M+H]⁺.

Example—114

N-(2-(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-ylamino)phenyl)propiolamide

The compound was synthesized following the approach outlined inProcedure 2C (Example 108) modifying step (g) to the followingprocedure: To a solution of1-[6-(2-amino-phenylamino)-pyrimidin-4-yl]-3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-methyl-urea(50 mg, 0.108 mmol) and DCC (46 mg, 0.22 mmol) in chloroform (50 mL) wasadded a solution of propiolic acid (16 mg, 0.22 mmol) in chloroform (50mL) at 0° C., and the resulting mixture was stirred at room temperatureovernight. Water (1 mL) was added to quench the reaction. The mixturewas concentrated and the residue was purified by reverse phase columnand prep-TLC to obtain the title compound (5 mg, yield: 9.1%). ¹H NMR(400 MHz, DMSO-d6) $12.04 (s, 1H), 10.36 (s, 1H), 9.03 (s, 1H), 8.45 (s,1H), 7.67 (d, 1H), 7.59 (d, 1H), 7.30-7.25 (m, 2H), 6.96 (s, 1H), 6.50(s, 1H), 4.42 (s, 1H), 4.00 (s, 6H), 3.35 (s, 3H); MS (ESI): 515 [M+H]⁺

Example—116

N-(2-{6-[3-(2,6-Dichloro-3,5-dimethoxy-phenyl)-1-(2-methoxy-ethyl)-ureido]-pyrimidin-4-ylamino}-phenyl)-acrylamide

The compound was synthesized following the approach outlined inProcedure 2G (Example 123), substituting1-(6-chloro-pyrimidin-4-yl)-3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-(2-methoxy-ethyl)-3-(2-trimethylsilanyl-ethoxymethyl)-urea(preparation shown below) in step (d) to afford the title compound (40mg, yield: 16% over five steps). ¹H NMR (400 MHz, DMSO-d6) δ 11.28 (s,1H), 9.71 (s, 1H), 8.85 (s, 1H), 8.38 (s, 1H), 7.70-7.68 (m, 1H),7.55-7.53 (m, 1H), 7.20-7.18 (m, 2H), 6.89 (s, 1H), 6.69 (s, 1H), 6.50(dd, 1H), 6.26 (d, 1H), 5.75 (d, 1H), 4.03 (t, 2H), 3.94 (s, 6H), 3.56(t, 2H), 3.24 (s, 3H); MS (ESI): 437 [M+H]⁺.

Preparation of1-(6-Chloro-pyrimidin-4-yl)-3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-(2-methoxy-ethyl)-3-(2-trimethylsilanyl-ethoxymethyl)-urea

a. (6-Chloro-pyrimidin-4-yl)-(2-methoxy-ethyl)-amine

To a solution of 4,6-dichloro-pyrimidine (2 g, 14 mmol) in iPrOH (70 mL)and DIPEA (1.94 g, 15 mmol) was added a solution of 2-methoxy-ethylamine(1.13 g, 15 mmol) at room temperature. The resulting mixture was stirredat room temperature for 1 hour. Water was added and the mixture wasextracted with DCM. The combined extracts were washed with brine, driedover anhydrous Na₂SO₄, and concentrated to give the crude product, whichwas purified by flash chromatography on silica to obtain the titlecompound (1.95 g, yield: 82%). MS (ESI): 188 [M+H]⁺.

b.1-(6-Chloro-pyrimidin-4-yl)-3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-(2-methoxy-ethyl)-3-(2-trimethylsilanyl-ethoxymethyl)-urea

To a solution of (6-chloro-pyrimidin-4-yl)-(2-methoxy-ethyl)-amine (300mg, 1.6 mmol) in DMF (10 mL) was added NaH (60%, 96 mg, 2.4 mmol) at 0°C., and the mixture was stirred for 10 minutes at room temperature. Asolution of 1-isocyanato-3,5-dimethoxy-benzene (590 mg, 2.4 mmol) in DMF(5 mL) was added dropwise at 0° C. The resulting mixture was stirred for30 minutes. SEMCl (400 mg, 2.4 mmol) in DMF (2 mL) was added and thereaction mixture was stirred at room temperature for 1 hour. Saturatedaqueous NH₄Cl was added to quench the reaction. The mixture was dilutedwith water and extracted with EtOAc. The combined extracts were washedwith water and brine, dried over anhydrous Na₂SO₄, and concentrated toafford the crude product, which was purified by flash chromatography onsilica to obtain the title compound (720 mg, yield: 78%). MS (ESI): 565[M+H]⁺.

Example—120

N-(2-{6-[3-(2,6-Dichloro-3,5-dimethoxy-phenyl)-1-(3-dimethylamino-propyl)-ureido]-pyrimidin-4-ylamino}-phenyl)-acrylamidetrifluoroacetic acid

The compound was synthesized following the approach outlined inProcedure 21 (Example 142), substituting 2-nitroaniline andN-(3-Dimethylamino-propyl)-N′-(2-nitro-phenyl)-pyrimidine-4,6-diamine(prepared by the method outlined below) in step (c) to afford the titlecompound (11 mg, yield: 7.5% over six steps). ¹H NMR (300 MHz, DMSO-d6)δ 11.30 (s, 1H), 9.80 (s, 1H), 9.28 (m, 1H), 8.92 (s, 1H), 8.42 (s, 1H),7.64 (d, 1H), 7.55 (d, 1H), 7.24-7.16 (m, 2H), 6.90 (s, 1H), 6.53 (s,1H), 6.49 (dd, 1H), 6.25 (d, 1H), 5.76 (d, 1H), 3.95 (s, 6H), 3.91-3.88(m, 2H), 3.11-3.05 (m, 2H), 2.74 (d, 6H), 1.97-1.92 (m, 2H); MS (ESI):588 [M+H]⁺.

Preparation ofN-(3-Dimethylamino-propyl)-N′-(2-nitro-phenyl)-pyrimidine-4,6-diamine

a. N′-(6-Chloro-pyrimidin-4-yl)-N,N-dimethyl-propane-1,3-diamine

To a solution of 4,6-dichloro-pyrimidine (1 g, 6.7 mmol) and DIPEA (1.03g, 8 mmol) in iPrOH (20 mL) was added N,N-dimethyl-propane-1,3-diamine(714 mg, 7 mmol) at room temperature. The resulting mixture was stirredfor 2 hours. Water was added and the mixture was extracted with DCM. Thecombined extracts were washed with brine, dried over anhydrous Na₂SO₄,and concentrated to give the crude product, which was purified by flashchromatography on silica to obtain the title compound (1.15 g, yield:80%). MS (ESI): 215 [M+H]⁺.

b. N-(3-Dimethylamino-propyl)-N′-(2-nitro-phenyl)-pyrimidine-4,6-diamine

A degassed mixture ofN′-(6-chloro-pyrimidin-4-yl)-N,N-dimethyl-propane-1,3-diamine (800 mg,3.74 mmol), nitro aniline (525 mg, 3.8 mmol), Pd₂(dba)₃ (348 mg, 0.38mmol), Xantphos (438 mg, 0.76 mmol) and Cs₂CO₃ (3.05 g, 9.35 mmol) intoluene (15 mL) was heated at 100° C. for 4 hours. The reaction wasconcentrated, and the residue was purified by reverse phasechromatography to obtain the title compound (530 mg, yield: 45%). MS(ESI): 317 [M+H]⁺.

Example—121

N-[2-(6-{3-(2,6-Dichloro-3,5-dimethoxy-phenyl)-1-[3-(4-methyl-piperazin-1-yl)-propyl]-ureido}-pyrimidin-4-ylamino)-phenyl]-acrylamide

The compound was synthesized following the approach outlined inProcedure 21 (Example 142), substituting 2-nitroaniline andN-[3-(4-Methyl-piperazin-1-yl)-propyl]-N′-(2-nitro-phenyl)-pyrimidine-4,6-diamine(preparation shown below) in step (e) to afford the title compound (8mg, yield: 1.2% over six steps). ¹H NMR (300 MHz, Methanol-d4) δ 8.17(s, 1H), 7.46 (d, 1H), 7.32 (d, 1H), 7.12-7.08 (m, 2H), 6.62 (s, 1H),6.29-6.14 (m, 3H), 5.58 (d, 1H), 3.82 (t, 2H), 3.75 (s, 6H), 2.27-2.19(m, 8H), 2.06 (s, 3H), 1.67 (t, 2H); MS (ESI): 643 [M+H]⁺.

Preparation ofN-[3-(4-Methyl-piperazin-1-yl)-propyl]-N′-(2-nitro-phenyl)-pyrimidine-4,6-diamine

a. (6-Chloro-pyrimidin-4-yl)-[3-(4-methyl-piperazin-1-yl)-propyl]-amine

To a solution of 4,6-dichloro-pyrimidine (1.5 g, 10 mmol) and DIPEA(1.55 g, 12 mmol) in iPrOH (50 mL) was added a solution of3-(4-methyl-piperazin-1-yl)-propylamine (1.73 g, 11 mmol) at roomtemperature. The resulting mixture was stirred at room temperature for 2hours. Water was added and the mixture was extracted with DCM. Thecombined extracts were washed with brine, dried over anhydrous Na₂SO₄,and concentrated to afford the title compound (1.4 g, 51%), which wasused directly in the next step without further purification. MS (ESI):270 [M+H]⁺.

b.N-[3-(4-Methyl-piperazin-J-yl)-propyl]-N′-(2-nitro-phenyl)-pyrimidine-4,6-diamine

A degassed mixture of(6-chloro-pyrimidin-4-yl)-[3-(4-methyl-piperazin-1-yl)-propyl]-amine(600 mg, 2.22 mmol), nitroanline (317 mg, 2.3 mmol), Pd₂(dba)₃ (210 mg,0.23 mmol), Xantphos (265 mg, 0.46 mmol) and Cs₂CO₃ (1.81 g, 5.55 mmol)in toluene (15 mL) was heated at 100° C. for 4 hours. The reaction wasconcentrated, and the residue was purified by reverse phasechromatography to obtain the title compound (400 mg, yield: 48%). MS(ESI): 372 [M+H]⁺.

Example—122

N-(2-{6-[3-(2,6-Dichloro-3,5-dimethoxy-phenyl)-1-methyl-ureido]-pyrimidin-4-ylamino}-5-piperidin-4-yl-phenyl)-acrylamidetrifluoroacetic acid

The compound was synthesized following the approach outlined inProcedure 2E (Example 111), replacing steps (e) and (f) with thefollowing procedure: To a solution of tert-butyl4-(3-amino-4-(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-((2-(trimethylsilyl)ethoxy)methyl)ureido)pyrimidin-4-ylamino)phenyl)piperidine-1-carboxylate(Procedure 2E, Example 111, 65 mg, 0.084 mmol) in THF (15 mL) was addeda solution of acryloyl chloride (10 mg/mL, 0.9 mL, 0.1 mmol) dropwise at−10° C., and the resulting mixture was stirred at 0° C. for 1 hour.

MeOH (5 mL) was added to quench the reaction, and the reaction wasconcentrated. The residue was dissolved in DCM (2 mL) and added dropwiseto a mixture of DCM/TFA (2:1, 3 mL). The mixture was stirred at roomtemperature for 1 hour and then concentrated. The residue was purifiedby prep-HPLC to afford the title compound (23 mg, yield: 47%). ¹H NMR(300 MHz, DMSO-d6) δ 11.92 (s, 1H), 9.64 (s, II), 8.98 (s, 1H), 8.60 (m,1H), 8.36 (s, 1H), 8.25 (min, 1H), 7.66 (s, 1H), 7.51 (d, 1H), 7.05 (d,1H), 6.90 (s, 1H), 6.52 (dd, 1H), 6.41 (s, 1H), 6.24 (d, 1H), 5.74 (d,1H), 3.96 (s, 6H), 3.49 (d, 2H), 3.28 (s, 3H), 3.02 (q, 2H), 2.85 (t,1H), 1.96 (d, 2H), 1.78 (q, 2H); MS (ESI): 600 [M+H]⁺.

Procedure 2G: Example—123

N-(2-{6-[3-(2,6-Dichloro-3,5-dimethoxy-phenyl)-1-methyl-ureido]-pyrimidin-4-ylamino}-5-morpholin-4-yl-phenyl)-acrylamide

a. di-tert-Butyl 4-bromo-2-nitrophenylcarbamate

A mixture of 4-bromo-2-nitroaniline (10 g, 46 mmol), (Boc)₂O (20.7 g, 95mmol) in THF (250 mL) was heated under reflux overnight. The mixture wasconcentrated to afford the title compound (19.2 g, yield: quant.) whichwas used directly in next step without further purification.

b. (4-Morpholin-4-yl-2-nitro-phenyl)-carbamic acid di-tert-butyl ester

A degassed mixture of tert-butyl 4-bromo-2-nitrophenylcarbamate (1 g,2.4 mmol), morpholine (314 mg, 3.6 mmol), Pd₂(dba)₃ (220 mg, 0.24 mmol),Xantphos (278 mg, 0.48 mmol) and Cs₂CO₃ (1.56 g, 4.8 mmol) in toluene(30 mL) was heated at 100° C. for 1 hour. The reaction was concentrated,and the residue was purified by flash chromatography on silica to obtaina crude mixture of the title compound and mono-Boc product (744 mg). Themixture was used directly in the next step without further purification.MS (ESI): 324 [M−Boc+H]⁺.

c. 4-Morpholin-4-yl-2-nitro-phenylamine

To a solution of (4-morpholin-4-yl-2-nitro-phenyl)-carbamic aciddi-tert-butyl ester and mono-Boc product (744 mg) in DCM (20 mL) wasadded TFA (10 mL) at 0° C., the resulting mixture was stirred for 3hours at room temperature. After removal of all volatiles in vacuo, theresidue was re-dissolved in DCM, neutralized by saturated aqueousNaHCO₃, and extracted with DCM.

The combined extracts were concentrated and the residue was purified byflash chromatography on silica to obtain the title compound (290 mg,yield: 54% over two steps). MS (ESI): 251 [M+H]⁺.

d.1-(2,6-Dichloro-3,5-dimethoxy-phenyl)-3-methyl-3-[6-(4-morpholin-4-yl-2-nitro-phenylamino)-pyrimidin-4-yl]-1-(2-trimethylsilanyl-ethoxymethyl)-urea

A degassed mixture of 4-morpholin-4-yl-2-nitro-phenylamine (290 mg, 1.3mmol),1-(6-chloro-pyrimidin-4-yl)-3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-methyl-3-(2-trimethylsilanyl-ethoxymethyl)-urea(Procedure 2E, step b; 624 mg, 1.2 mmol), Pd₂(dba)₃ (110 mg, 0.12 mmol),Xantphos (139 mg, 0.24 mmol) and Cs₂CO₃ (782 mg, 2.4 mmol) in toluene(15 mL) was heated at 100° C. for 2.5 hours. The reaction wasconcentrated, and the residue was purified by chromatography flash onsilica to obtain the title compound (440 mg, yield: 49%) as a red solid.MS (ESI): 708 [M+H]⁺.

e.{6-[3-(2,6-Dichloro-3,5-dimethoxy-phenyl)-1-methyl-3-(2-trimethylsilanyl-ethoxymethyl)-ureido]-pyrimidin-4-yl}-(4-morpholin-4-yl-2-nitro-phenyl)-carbamicacid tert-butyl ester

A mixture of1-(2,6-dichloro-3,5-dimethoxy-phenyl)-3-methyl-3-[6-(4-morpholin-4-yl-2-nitro-phenylamino)-pyrimidin-4-yl]-1-(2-trimethylsilanyl-ethoxymethyl)-urea(200 mg, 0.28 mmol), (Boc)₂O (93 mg, 0.42 mmol) and catalytic amount ofDMAP in THF (10 mL) was heated under reflux for 1 hour. The mixture wasconcentrated and the residue was used for the next step without furtherpurification. MS (ESI): 808 [M+H]⁺.

f.(2-Amino-4-morpholin-4-yl-phenyl)-{6-[3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-methyl-3-(2-trimethylsilanyl-ethoxymethyl)-ureido]-pyrimidin-4-yl}-carbamicacid tert-butyl ester

To a solution of{6-[3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-methyl-3-(2-trimethylsilanyl-ethoxymethyl)-ureido]-pyrimidin-4-yl}-(4-morpholin-4-yl-2-nitro-phenyl)-carbamicacid tert-butyl ester (crude, prepared above) in MeOH (20 mL) was addedRaney-Ni (suspension in water) at room temperature, the resultingmixture was stirred for 2 hours under hydrogen atmosphere (1 atm). Thereaction was filtered and concentrated. The residue was washed twicewith MeOH to obtain title product (160 mg, yield: 70%). MS (ESI):778[M+H]⁺.

g.(2-Acryloylamino-4-morpholin-4-yl-phenyl)-{6-[3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-(2-trimethylsilanyl-ethoxymethyl)-ureido]-pyrimidin-4-yl}-carbamicacid tert-butyl ester

To a solution of(2-amino-4-morpholin-4-yl-phenyl)-{6-[3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-methyl-3-(2-trimethylsilanyl-ethoxymethyl)-ureido]-pyrimidin-4-yl}-carbamicacid tert-butyl ester (80 mg, 0.103 mmol) in DCM (5 mL) was added asolution of TEA (10 mg/mL, 1.2 mL, 0.12 mmol) and a solution of acryloylchloride (10 mg/mL, 1 mL, 0.11 mmol) dropwise at 0° C., and theresulting mixture was stirred at room temperature for 1 hour. LC-MSshowed that the reaction was complete. Water (5 mL) was added to quenchthe reaction, and the reaction mixture was extracted with DCM. Thecombined extracts were washed with brine, dried over anhydrous sodiumsulfate and filtered. The filtrate was evaporated under vacuum to givecrude product, which was used for the next step without furtherpurification.

h.N-(2-{6-[3-(2,6-Dichloro-3,5-dimethoxy-phenyl)-1-methyl-ureido]-pyrimidin-4-ylamino}-5-morpholin-4-yl-phenyl)-acrylamide

To a solution of(2-acryloylamino-4-morpholin-4-yl-phenyl)-{6-[3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-methyl-3-(2-trimethylsilanyl-ethoxymethyl)-ureido]-pyrimidin-4-yl}-carbamicacid tert-butyl ester (crude, prepared above) in CH₂Cl₂ (10 mL) wasadded TFA (3 mL) at 0° C., the resulting mixture was stirred for 1 hourat room temperature. The reaction mixture was concentrated andneutralized with NH₃.H₂O to give the crude compound, which was purifiedby Prep-HPLC to obtain title compound (10 mg, yield: 16% in two steps).¹H NMR (300 MHz, DMSO-d6) δ 12.05 (s, 1H), 9.60 (s, 1H), 8.74 (s, 1H),8.32 (s, 1H), 7.33-7.30 (m, 2H), 6.89 (s, 1H), 6.83 (d, 1H), 6.48 (dd,1H), 6.25-6.21 (m, 2H), 5.72 (d, 1H), 3.96 (s, 6H), 3.73 (br, 4H), 3.44(s, 3H), 3.10 (br, 4H); MS (ESI): 602 [M+H]⁺

Example—124

N-[2-{6-[3-(2,6-Dichloro-3,5-dimethoxy-phenyl)-1-methyl-ureido]-pyrizmidin-4-ylamino}-5-(3,5-dimethyl-piperazin-1-yl)-phenyl]-acrylamide

The compound was synthesized following the approach outlined inProcedure 2G (Example 123), substituting4-(3,5-dimethyl-piperazin-1-yl)-2-nitro-phenylamine (preparation shownbelow) in step (d) to afford the title compound (26 mg, yield: 30% overthree steps). ¹H NMR (300 MHz, Methanol-d4) δ 8.32 (s, 1H), 7.36-7.33(m, 2H), 6.95 (d, 1H), 6.80 (s, 1H), 6.43-6.37 (m, 2H), 6.15 (s, 1H),5.77 (d, 1H), 3.95 (s, 6H), 3.70 (d, 2H), 3.29 (s, 3H), 3.18-3.13 (m,2H), 2.45 (t, 2H), 1.24 (d, 6H); MS (ESI): 629 [M+H]⁺.

Preparation of 4-(3,5-Dimethyl-piperazin-1-yl)-2-nitro-phenylamine

a. di-tert-Butyl 4-bromo-2-nitrophenylcarbamate

A mixture of 4-bromo-2-nitroaniline (10 g, 46 mmol), (Boc)₂O (20.7 g, 95mmol) in THF (250 mL) was heated under reflux overnight. The mixture wasconcentrated to afford the title compound (19.2 g, quant) which was usedfor the next step without further purification.

b. [4-(3,5-Dimethyl-piperazin-1-yl)-2-nitro-phenyl]-carbamic aciddi-tert-butyl ester

A degassed mixture of di-tert-Butyl 4-bromo-2-nitrophenylcarbamate (1 g,2.4 mmol), 2,6-dimethyl-piperazine (410 mg, 3.6 mmol), Pd₂(dba)₃ (220mg, 0.24 mmol), Xantphos (278 mg, 0.48 mmol) and Cs₂CO₃ (1.56 g, 4.8mmol) in toluene (30 mL) was heated at 100° C. for 1 hour. The reactionwas concentrated, and the residue was purified by flash chromatographyon silica to obtain a mixture of the title compound and mono-Boc product(600 mg). The mixture was used directly in the next step without furtherpurification. MS (ESI): 350 [M−Boc+H]⁺.

c. 4-(3,5-Dimethyl-piperazin-1-yl)-2-nitro-phenylamine

To a solution of[4-(3,5-dimethyl-piperazin-1-yl)-2-nitro-phenyl]-carbamic aciddi-tert-butyl ester and mono-Boc product (600 mg) in DCM (20 mL) wasadded TFA (10 mL) at 0° C., the resulting mixture was stirred for 3hours at room temperature. After removal of all volatiles in vacuo, theresidue was re-dissolved in DCM, neutralized by saturated aqueousNaHCO₃, and extracted with DCM. The combined extracts were concentratedand the residue was purified by flash chromatography on silica to obtainthe title compound (233 mg, yield: 39% over two steps). MS (ESI): 251[M+H]⁺.

Example—125

N-[2-{6-[3-(2,6-Dichloro-3,5-dimethoxy-phenyl)-1-methyl-ureido]-pyrimidin-4-ylamino}-5-(4-methanesulfonyl-piperazin-1-yl)-phenyl]-acrylamide

The compound was synthesized following the approach outlined inProcedure 2G (Example 123), substituting4-(4-methanesulfonyl-piperazin-1-yl)-2-nitro-phenylamine (preparationshown below) in step (d) to afford the title compound (15 mg, yield:9.6% in five steps). ¹H NMR (300 MHz, DMSO-d6) δ 12.03 (s, 1H), 9.62 (s,1H), 8.76 (s, 1H), 8.33 (s, 1H), 7.37-7.32 (m, 2H), 6.89-6.83 (m, 2H),6.46 (dd, 1H), 6.26-6.21 (m, 2H), 5.72 (d, 1H), 3.93 (s, 6H), 3.34-3.16(m, 11H), 2.93 (s, 3H); MS (ESI): 679 [M+H]⁺.

Preparation of 4-(4-Methanesulfonyl-piperazin-1-yl)-2-nitro-phenylamine

a. di-tert-Butyl 4-bromo-2-nitrophenylcarbamate

A mixture of 4-bromo-2-nitroaniline (10 g, 46 mmol), (Boc)₂O (20.7 g, 95mmol) in THF (250 mL) was heated under reflux overnight. The mixture wasconcentrated to afford the title compound (19.2 g, quant) which was useddirectly in next step without further purification.

b. [4-(4-Methanesulfonyl-piperazin-1-yl)-2-nitro-phenyl]-carbamic aciddi-tert-butyl ester

A degassed mixture of di-tert-butyl 4-bromo-2-nitrophenylcarbamate (1 g,2.4 mmol), 1-methanesulfonyl-piperazine (590 mg, 3.6 mmol), Pd₂(dba)₃(220 mg, 0.24 mmol), Xantphos (278 mg, 0.48 mmol) and Cs₂CO₃ (1.56 g,4.8 mmol) in toluene (30 mL) was heated at 100° C. for 1 hour. Thereaction was concentrated, and the residue was purified by flashchromatography on silica to obtain a mixture of the title compound andmono-Boc product (755 mg). The mixture was used directly in next stepwithout further purification. MS (ESI): 400 [M−Boc+H]⁺.

c. 4-(4-Methanesulfonyl-piperazin-1-yl)-2-nitro-phenylamine

To a solution of[4-(4-methanesulfonyl-piperazin-1-yl)-2-nitro-phenyl]-carbamic aciddi-tert-butyl ester and mono-Boc product (755 mg) in DCM (20 mL) wasadded TFA (10 mL) at 0° C., the resulting mixture was stirred for 3hours at RT, After removal of all volatiles in vacuo, the residue wasre-dissolved in DCM, neutralized by saturated aqueous NaHCO₃, andextracted with DCM. The combined extracts were concentrated and theresidue was purified by flash chromatography on silica to obtain thetitle compound (290 mg, yield: 40% in two steps). MS (ESI): 301 [M+H]⁺.

Example—126

N-[2-{6-[3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-methyl-ureido]-pyrimidin-4-ylamino}-5-(3-dimethylamino-propyl)-phenyl]-acrylamide

The compound was synthesized following the approach outlined inProcedure 2G (Example 123), substituting4-(3-Dimethylamino-prop-1-ynyl)-2-nitro-phenylamine (preparation shownbelow) in step (d) and platinum oxide in step (f) to afford the titlecompound (4 mg, yield: 1.4% in five steps). ¹H-NMR (300 MHz, DMSO-d6) δ11.98 (s, 1H), 9.66 (d, 1H), 8.88 (s, 1H), 8.36 (s, 1H), 7.52 (d, 1H),7.44 (d, 1H), 7.04 (dd, 1H), 6.90 (s, 1H), 6.48 (dd, 1H), 6.34 (s, 1H),6.24 (dd, 1H), 5.72 (dd, 1H), 3.94 (s, 6H), 3.25 (s, 3H), 2.59 (t, 2H),2.24 (t, 2H), 2.11 (s, 6H), 1.71 (m, 2H); MS (ESI) 602 [M+H]⁺.

Preparation of 4-(3-Dimethylamino-prop-1-ynyl)-2-nitro-phenylamine

a. 4-(3-Dimethylamino-prop-1-ynyl)-2-nitro-phenylamine

To a stirred solution of 4-bromo-2-nitro-phenylamine (1.08 g, 5 mmol)and dimethyl-prop-2-ynyl-amine (1.0 g, 12 mmol) in TEA (20 mL) was addedPd(PPh₃)₂Cl₂ (0.7 g, 1 mmol) and CuI (360 mg, 2 mmol). The solution wasstirred at 60° C. under nitrogen for 3 hours. The solution wasevaporated with silica gel and purified by flash chromatography onsilica to give the title compound (1.1 g, 70% purity). The titlecompound was taken directly to the next step without furtherpurification. MS (ESI) 221 [M+H]⁺.

Example—127

N-[2-{6-[3-(2,6-Dichloro-3,5-dimethoxy-phenyl)-1-methyl-ureido]-pyrimidin-4-ylamino}-5-(2-dimethylamino-ethoxy)-phenyl]-acrylamide

The compound was synthesized following the approach outlined inProcedure 2G for compound 123, substituting4-(2-Dimethylamino-ethoxy)-2-nitro-phenylamine (preparation shown below)in step (d) and platinum oxide in step (f) to afford the title compound(4.3 mg, yield: 8% over five steps). ¹H-NMR (300 MHz, DMSO-d6) δ 12.04(s, 1H), 9.58 (s, 1H), 8.81 (s, 1H), 8.33 (s, 1H), 7.43 (d, 1H), 7.33(d, 1H), 6.89 (s, 1H), 6.79 (dd, 1H), 6.51 (dd, 1H), 6.26-6.20 (m, 2H),5.73 (dd, 1H), 4.04 (t, 2H), 3.96 (s, 6H), 3.27 (s, 3H), 2.67 (t, 2H),2.24 (s, 6H); MS (ESI) 604 [M+H]⁺.

Preparation of 4-(2-Dimethylamino-ethoxy)-2-nitro-phenylamine

a. 4-(2-Dimethylamino-ethoxy)-2-nitro-phenylamine

To a stirred solution of 4-amino-3-nitro-phenol (1.54 g, 10 mmol) and(2-chloro-ethyl)-dimethyl-amine hydrogen chloride (1.43 g, 10 mmol) inbutanone (40 mL) was added Cs₂CO₃ (10 g, 30 mmol) and NaI (150 mg, 1mmol). The solution was slowly heated to 80° C. over one hour. Then thesolution was stirred at 80° C. for 2 hours. The solution was filteredthrough Celite® and washed with acetone. The solution was evaporatedwith silica gel and purified by flash chromatography on silica to affordthe title compound (1.0 g, yield: 45%) as a brown solid. ¹H-NMR (300MHz, DMSO-d6) δ 7.38 (d, 1H), 7.24 (s, 2H), 7.16 (dd, 2H), 6.99 (d, 1H),3.98 (t, 2H), 2.58 (t, 2H), 2.20 (s, 6H); MS (ESI) 226 [M+H]⁺.

Example—128

But-2-ynoic acid(2-{6-[3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-methyl-ureido]-pyrimidin-4-ylamino}-phenyl)-amide

The compound was synthesized following the approach outlined inProcedure 2G (Example 123), substituting 2-nitro-phenylamine in step(d), omitting step (e), and replacing step (g) with the followingprocedure below to afford the title compound (7 mg, yield: 4.8% overfive steps). ¹H NMR (300 MHz, DMSO-d6) δ 11.97 (s, 1H), 10.07 (s, 1H),8.89 (s, 1H), 8.39 (s, 1H), 7.60 (d, 1H), 7.53 (d, 1H), 7.24-7.15 (m,2H), 6.90 (s, 1H), 6.44 (s, 1H), 3.93 (s, 6H), 3.29 (s, 3H), 2.22 (s,3H); MS (ESI): 529 [M+H]⁺.

To a solution of(2-amino-phenyl)-{6-[3-(2,65-dichloro-3,5-dimethoxy-phenyl)-1-methyl-3-(2-trimethylsilanyl-ethoxymethyl)-ureido]-pyrimidin-4-yl}-carbamicacid tert-butyl ester (50 mg, 0.075 mmol) and DCC (42 mg, 0.2 mmol) inDCM (50 mL) was added a solution of but-2-ynoic acid (13 mg, 0.15 mmol)in DCM (10 mL) at 0° C., and the resulting mixture was stirred at roomtemperature overnight. Water (1 mL) was added to quench the reaction.The mixture was concentrated and the residue was purified by reversephase chromatography to obtain the title compound (20 mg, yield: 34%).MS (ESI): 659 [M+H]⁺.

Compound 129 was synthesized in a similar manner as compound 100.

Example—130

N-(2-{6-[1-Benzyl-3-(2,6-dichloro-3,5-dimethoxy-phenyl)-ureido]-pyrimidin-4-ylamino}-phenyl)-acrylamide

The compound was synthesized following the approach outlined inProcedure 2G (Example 123), substituting 2-nitro-phenylamine and1-benzyl-1-(6-chloro-pyrimidin-4-yl)-3-(2,6-dichloro-3,5-dimethoxy-phenyl)-3-(2-trimethylsilanyl-ethoxymethyl)-urea(preparation shown below) in step (d) to afford the title compound (26mg, yield: 20% over five steps). ¹H NMR (300 MHz, DMSO-d6) δ 12.27 (s,1H), 9.68 (s, 1H), 8.90 (s, 1H), 8.39 (s, 1H), 7.69 (d, 1H), 7.33-7.19(m, 6H), 6.92 (s, 1H), 6.45 (dd, 1H), 6.24 (d, 1H), 6.18 (s, 1H), 5.74(d, 1H), 5.07 (s, 2H), 3.95 (s, 6H); MS (ESI): 593 [M+H]⁺.

Preparation ofI-Benzyl-1-(6-chloro-pyrimidin-4-yl)-3-(2,6-dichloro-3,5-dimethoxy-phenyl)-3-(2-trimethylsilanyl-ethoxymethyl)-urea

a. Benzyl-(6-chloro-pyrimidin-4-yl)-amine

To a solution of 4,6-dichloro-pyrimidine (1.5 g, 10 mmol) in iPrOH (40mL) and DIPEA (1.55 g, 12 mmol) was added a solution of benzylamine(1.28 g, 12 mmol) at room temperature. The resulting mixture was stirredat room temperature for 2.5 hours. Water was added and the mixture wasextracted with DCM. The combined extracts were washed with brine, driedover anhydrous Na₂SO₄, and concentrated to give the crude product, whichwas purified by flash chromatography on silica to obtain the titlecompound (1.5 g, yield: 68%) as a white solid. ¹H NMR (300 MHz, CDCl₃) δ8.35 (br s, 1H), 7.20-7.38 (m, 5H), 6.35 (s, 1H), 4.52 (s, 2H); MS(ESI): 220 [M+H]⁺.

b.J-Benzyl-1-(6-chloro-pyrimidin-4-yl)-3-(2,6-dichloro-3,5-dimethoxy-phenyl)-3-(2-trimethylsilanyl-ethoxymethyl)-urea

To a solution of benzyl-(6-chloro-pyrimidin-4-yl)-amine (800 mg, 3.64mmol) in DMF (15 mL) was added NaH (60%, 218 mg, 5.45 mmol) at 0° C.,and the mixture was stirred for 10 minutes at room temperature. Asolution of 1-isocyanato-3,5-dimethoxy-benzene (Procedure 2A, steps a-d;1.35 g, 5.45 mmol) in DMF (2 mL) was added dropwise at 0° C. Theresulting mixture was stirred at room temperature for 2 hours. Saturatedaqueous NH₄Cl (2 mL) was added to quench the reaction. The mixture wasconcentrated and extracted with DCM. The combined extracts were washedwith brine, dried over anhydrous Na₂SO₄, and concentrated to give thecrude product, which was purified by flash chromatography on silica toobtain the title compound (1.7 g, yield: 77%). MS (ESI): 599 [M+H]⁺.

Example—131

N-(2-{6-[3-(2,6-Dichloro-3,5-dimethoxy-phenyl)-1-pyridin-2-ylmethyl-ureido]-pyrimidin-4-ylamino}-phenyl)-acrylamide

The compound was synthesized following the approach outlined inProcedure 2G (Example 123), substituting 2-nitro-phenylamine and1-(6-chloro-pyrimidin-4-yl)-3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-pyridin-2-ylmethyl-3-(2-trimethylsilanyl-ethoxymethyl)-urea(preparation shown below) in step (d) to afford the title compound (35mg, yield: 13% over five steps). ¹H NMR (300 MHz, DMSO-d6) δ 12.15 (s,1H), 9.65 (s, 1H), 8.83 (s, 1H), 8.50 (s, 1H), 8.39 (s, 1H), 7.80 (t,1H), 7.66 (d, 1H), 7.31-7.25 (m, 3H), 7.17 (t, 1H), 7.09 (t, 1H), 6.91(s, 11H), 6.45 (dd, 1H), 6.39 (s, 1H), 6.23 (d, 1H), 5.74 (d, 1H), 5.17(s, 2H), 3.94 (s, 6H); MS (ESI): 594 [M+H]⁺.

Preparation of1-(6-Chloro-pyrimidin-4-yl)-3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-pyridin-2-ylmethyl-3-(2-trimethylsilanyl-ethoxymethyl)-urea

a. (6-Chloro-pyrimidin-4-yl)-pyridin-2-ylmethyl-amine

To a solution of 4,6-dichloro-pyrimidine (1 g, 7 mmol) in iPrOH (40 mL)and DIPEA (1.16 g, 9 mmol) was added a solution of2-pyridinylmethanamine (970 mg, 9 mmol) at room temperature. Theresulting mixture was stirred at room temperature for 2.5 hours. Waterwas added and the mixture was extracted with DCM. The combined extractswere washed with brine, dried over anhydrous sodium sulfate andconcentrated to give a crude product, which was purified by flashchromatography on silica to obtain the title compound (1.2 g, yield:78%). MS (ESI): 221 [M+H]⁺.

b.1-(6-Chloro-pyrimidin-4-yl)-3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-pyridin-2-ylmethyl-3-(2-trimethylsilanyl-ethoxymethyl)-urea

To a solution of (6-chloro-pyrimidin-4-yl)-pyridin-2-ylmethyl-amine (200mg, 0.91 mmol) in DMF (5 mL) was added NaH (60%, 55 mg, 1.37 mmol) at 0°C., and the mixture was stirred for 10 minutes at room temperature. Asolution of 1-isocyanato-3,5-dimethoxy-benzene (Procedure 2A, a-d; 337mg, 1.37 mmol) in DMF (2 mL) was added dropwise at 0° C. The resultingmixture was stirred for 30 minutes. SEMCl (230 mg, 1.37 mmol) in DMF (2mL) was added and the reaction mixture was stirred at room temperaturefor 1 hour. Saturated aqueous NH₄Cl was added to quench the reaction.The mixture was diluted with water and extracted with EtOAc. Thecombined extracts were washed with water and brine, dried over anhydrousNa₂SO₄ and filtered. The filtrate was concentrated to give a crudeproduct, which was purified by flash chromatography on silica to obtainthe title product (420 mg, yield: 78%). MS (ESI): 598 [M+H]⁺.

Example—132

N-(2-{6-[3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-pyridin-4-ylmethyl-ureido]-pyrimidin-4-ylamino}-phenyl)-acrylamide

The compound was synthesized following the approach outlined inProcedure 2G (Example 123), substituting3-(2,6-Dichloro-3,5-dimethoxy-phenyl)-1-[6-(2-nitro-phenylamino)-pyrimidin-4-yl]-1-pyridin-4-ylmethyl-ureain step (e) to afford the title compound (14 mg, yield: 8.1%). ¹H NMR(300 MHz, DMSO-d6) δ 12.03 (s, 1H), 9.68 (s, 1H), 8.92 (s, 1H), 8.49 (d,2H), 8.39 (s, 1H), 7.67 (d, 1H), 7.27-7.17 (m, 4H), 7.10 (t, 1H), 6.92(s, 1H), 6.48 (dd, 1H), 6.24 (d, 1H), 6.15 (s, 1H), 5.74 (d, 1H), 5.09(s, 2H), 3.94 (s, 6H); MS (ESI): 594 [M+H]⁺.

Preparation of3-(2,6-Dichloro-3,5-dimethoxy-phenyl)-1-[6-(2-nitro-phenylamino)-pyrimidin-4-yl]-1-pyridin-4-ylmethyl-urea

a. (6-Chloro-pyrimidin-4-yl)-pyridin-4-ylmethyl-amine

To a solution of 4,6-dichloro-pyrimidine (1.5 g, 10.5 mmol) and DIPEA(1.62 g, 12.6 mmol) in iPrOH (40 mL) was added 4-pyridinylmethanamine(1.2 g, 11 mmol) at room temperature. The resulting mixture was stirredat room temperature for 2 hours. Water was added and the mixture wasextracted with DCM. The combined extracts were washed with brine, driedover anhydrous Na₂SO₄, and concentrated to give the crude product, whichwas purified by flash chromatography on silica to obtain the titlecompound (1.8 g, yield: 80%). MS (ESI): 221 [M+H]⁺.

b. N-(2-Nitro-phenyl)-N pyridin-4-ylmethyl-pyrimidine-4,6-diamine

A degassed mixture of (6-chloro-pyrimidin-4-yl)-pyridin-4-ylmethyl-amine(500 mg, 2.27 mmol), 2-nitroaniline (317 mg, 2.3 mmol), Pd₂(dba)₃ (200mg, 0.22 mmol), Xantphos (253 mg, 0.44 mmol) and Cs₂CO₃ (1.48 g, 9.35mmol) in toluene (10 mL) was heated at 100° C. for 3 hours. The reactionwas concentrated, and the residue was purified by reverse phasechromatography followed by flash chromatography on silica to obtain thetitle compound (330 mg, yield: 45%). MS (ESI): 323 [M+H]⁺.

c.3-(2,6-Dichloro-3,5-dimethoxy-phenyl)-1-[6-(2-nitro-phenylamino)-pyrimidin-4-yl]-1-pyridin-4-ylmethyl-urea

To a solution ofN-(2-nitro-phenyl)-N′-pyridin-4-ylmethyl-pyrimidine-4,6-diamine (330 mg,1.02 mmol) in DMF (10 mL) was added NaH (60%, 56 mg, 1.4 mmol) at 0° C.,and the mixture was stirred for 30 minutes at room temperature. Asolution of 1-isocyanato-3,5-dimethoxy-benzene (Procedure 2A, steps a-d;345 mg, 1.4 mmol) in DMF (2 mL) was added dropwise at 0° C. Theresulting mixture was stirred at room temperature for 2 hours. Saturatedaqueous NH₄Cl (2 mL) was added to quench the reaction. The mixture wasconcentrated and extracted with DCM. The combined extracts were washedwith brine, dried over anhydrous Na₂SO₄, and concentrated to give thecrude product, which was purified by prep-TLC to obtain the titlecompound (190 mg, yield: 33%). MS (ESI): 570 [M+H]⁺.

Example—133

Preparation of 5-(4-ethyl-piperazin-1-yl)-2-nitro-phenylamine

1. 5-(4-Ethyl-piperazin-1-yl)-2-nitro-phenylamine

A mixture of 1-ethyl-piperazine (1.2 mL, 9.6 mmol),5-fluoro-2-nitro-phenylamine (1 g, 6.4 mmol), DIPEA (1.24 g, 9.6 mmol)in DMF (15 mL) was heated at 80° C. overnight. The reaction mixture waspoured into ice water and extracted with EtOAc. The combined extract waswashed with brine, dried over anhydrous Na₂SO₄, and concentrated toobtain a crude product, which was purified by flash chromatography onsilica to afford the title compound (1 g, yield: 63%). ESI-MS: 251[M+H]⁺.

N-[2-{6-[3-(2,6-Dichloro-3,5-dimethoxy-phenyl)-1-methyl-ureido]-pyrimidin-4-ylamino}-4-(4-ethyl-piperazin-1-yl)-phenyl]-acrylamidetrifluoroacetic acid salt

The compound was synthesized following the approach outlined inProcedure 2G (Example 123), substituting5-(4-ethyl-piperazin-1-yl)-2-nitro-phenylamine in step (a) to afford thetitle compound (20 mg, yield: 39%). ¹H NMR (300 MHz, Methanol-d4) δ 8.37(s, 1H), 7.49 (d, 1H), 7.25 (s, 1H), 6.98 (d, 1H), 6.82 (s, 1H),6.43-6.38 (m, 3H), 5.78 (d, 1H), 3.96-3.88 (m, 8H), 3.68-3.64 (m, 2H),3.37 (s, 3H), 3.33-3.08 (m, 6H), 1.40 (t, 3H); MS (ESI): 629 [M+H]⁺.

Example—135

N-[2-{6-[3-(2,6-Dichloro-3,5-dimethoxy-phenyl)-1-methyl-ureido]-pyrimidin-4-ylamino}-5-(4-ethyl-piperazin-1-ylmethyl)-phenyl]-acrylamide

The compound was synthesized following the approach outlined inProcedure 2G (Example 123), substituting4-(4-Ethyl-piperazin-1-ylmethyl)-2-nitro-phenylamine (preparation shownbelow) in step (d) to afford the title compound (30 mg, yield: 31% overfive steps). ¹H NMR (300 MHz, DMSO-d6) δ 11.97 (s, 1H), 9.71 (s, 1H),8.95 (s, 1H), 8.37 (s, 1H), 7.60 (s, 1H), 7.50 (d, 1H), 7.12 (d, 1H),6.90 (s, 1H), 6.50 (dd, 1H), 6.39 (s, 1H), 6.25 (d, 1H), 5.73 (d, 1H),3.93 (s, 6H), 3.48 (s, 2H), 3.32 (s, 3H), 2.50-2.25 (m, 10H), 0.98 (t,3H); MS (ESI): 643 [M+H]⁺.

Preparation of 4-(4-Ethyl-piperazin-1-ylmethyl)-2-nitro-phenylamine

a. 4-(4-Ethyl-piperazin-1-ylmethyl)-2-nitro-phenylamine

To a stirred solution of 1-ethyl-piperazine (1.37 g, 12 mmol) in MeOH(30 mL) was added Ti(O^(i)Pr)₄ (1.73 g, 6 mmol). Then the solution wasstirred at room temperature for 15 min. Then4-amino-3-nitro-benzaldehyde (Procedure 2F, steps a-b, 1.5 g, 9 mmol) inMeOH (10 mL) was added and the solution was stirred at room temperatureovernight. Then NaBH₄ (380 mg, 10 mmol) was added and the solution wasstirred at room temperature for 1 hour. The solution was diluted withEtOAc and filtered. The filtrate was washed with water and brine, driedover anhydrous Na₂SO₄. Concentration gave product, which was purified byflash chromatography on silica to obtain the title compound (800 mg,yield: 34%). MS (ESI): 265 [M+H]⁺

Example—136

N-(2-{6-[3-(2,6-Dichloro-3,5-dimethoxy-phenyl)-1-(3-methyl-isoxazol-5-ylmethyl)-ureido]-pyrimidin-4-ylamino}-phenyl)-acrylamide

The compound was synthesized following the approach outlined inProcedure 2G (Example 123), substituting1-(6-Chloro-pyrimidin-4-yl)-3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-(3-methyl-isoxazol-5-ylmethyl)-3-(2-trimethylsilanyl-ethoxymethyl)-urea(preparation shown below) in step (d) and iron/acetic acid at 60° C. instep (f) to afford the title compound (26 mg, yield: 12% in five steps).¹H NMR (300 MHz, DMSO-d6) δ 11.78 (s, 1H), 9.72 (s, 1H), 8.97 (s, 1H),8.40 (s, 1H), 7.66 (d, 1H), 7.45 (d, 1H), 7.21-7.15 (m, 2H), 6.90 (s,1H), 6.52-6.43 (m, 2H), 6.26 (d, 1H), 6.09 (s, 1H), 5.74 (d, 1H), 5.16(s, 2H), 3.93 (s, 6H), 2.18 (s, 3H); MS (ESI): 598 [M+H]⁺

Preparation of1-(6-Chloro-pyrimidin-4-yl)-3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-(3-methyl-isoxazol-5-ylmethyl)-3-(2-trimethylsilanyl-ethoxymethyl)-urea

a. (6-Chloro-pyrimidin-4-yl)-(3-methyl-isoxazol-5-ylmethyl)-amine

To a solution of 4,6-dichloro-pyrimidine (660 mg, 4.46 mmol) in iPrOH(40 mL) and DIEA (690 mg, 5.35 mmol) was added a solution ofC-pyridin-2-yl-methylamine (560 mg, 5 mmol) at room temperature. Theresulting mixture was stirred at room temperature for 2 hours. Water wasadded and the mixture was extracted with DCM. The combined extracts werewashed with brine, dried over anhydrous sodium sulfate, andconcentration under vacuum to give the crude product. The crude productwas purified by flash chromatography on silica to obtain the titlecompound (650 mg, yield: 65%). MS (ESI): 225 [M+H]⁺.

b.1-(6-Chloro-pyrimidin-4-yl)-3-(2,6-dichloro-3,5-dimethoxy-phenyl)-1-(3-methyl-isoxazol-5-ylmethyl)-3-(2-trimethylsilanyl-ethoxymethyl)-urea

To a solution of(6-chloro-pyrimidin-4-yl)-(3-methyl-isoxazol-5-ylmethyl)-amine (300 mg,1.34 mmol) in DMF (5 mL) was added NaH (60%, 80 mg, 2 mmol) at 0° C.,and the mixture was stirred for 10 minutes at room temperature. Asolution of 1-isocyanato-3,5-dimethoxy-benzene (Procedure 2A, steps a-d,337 mg, 1.37 mmol) in DMF (2 mL) was added dropwise at 0° C. Theresulting mixture was stirred for 0.5 hour. SEMCl (230 mg, 1.37 mmol) inDMF (2 mL) was added. The reaction mixture was stirred at roomtemperature for 1 hour. Sat. aqueous NH₄Cl was added to quench thereaction. The mixture was diluted with water and extracted with EtOAc.The combined extracts were washed with water and brine, dried overanhydrous sodium sulfate and filtered. The filtrate was evaporated undervacuum to give the crude product, which was purified by flashchromatography on silica to obtain the title product (440 mg, yield:55%). MS (ESI): 604 [M+H]⁺

Example—137

N-[2-{6-[3-(2,6-Dichloro-3,5-dimethoxy-phenyl)-1-methyl-ureido]-pyrimidin-4-ylamino}-4-(2-dimethylamino-ethyl)-phenyl]-acrylamideTFA salt

The compound was synthesized following the approach outlined inProcedure 2G substituting 5-(2-dimethylamino-ethyl)-2-nitro-phenylamine(prepared by the method outlined below) in step (d) to afford the titlecompound (39 mg, yield: 29%) as a TFA salt. ¹H NMR (300 MHz, DMSO-d6) δ11.86 (s, 1H), 9.76-9.70 (m, 2H), 9.02 (s, 1H), 8.40 (s, 1H), 7.86 (d,1H), 7.51 (s, 1H), 7.12 (d, 1H), 6.90 (s, 1H), 6.57-6.48 (m, 2H), 6.24(d, 1H), 5.74 (d, 1H), 3.94 (s, 6H), 3.35-3.28 (m, 5H), 2.98-2.92 (m,2H), 2.79 (s, 6H); MS (ESI): 588 [M+H]⁺.

Preparation of 5-(2-Dimethylamino-ethyl)-2-nitro-phenylamine

a. (2-(3-Fluoro-4-nitro-phenyl)-vinyl-dimethyl-amine

A mixture of 2-fluoro-4-methyl-1-nitro-benzene (3 g, 19.3 mmol),N,N-dimethylformamide dimethylacetal (10 mL) and 3 mL of DMF (30 mL) washeated at 125° C. for 1 hour. The mixture was cooled and concentratedunder reduced pressure to give a purple solid. Trituration with hexanesgave the pure title product (2.5 g, yield: 63%). MS (ESI): 211 [M+H]⁺.

b. [2-(3-Fluoro-4-nitro-phenyl)-ethyl]-dimethyl-amine

To a solution of [2-(3-fluoro-4-nitro-phenyl)-vinyl]-dimethyl-amine (1.7g, 8 mmol) in MeOH, was added NaBH₃CN (770 mg, 12 mmol) and one drop ofAcOH. The reaction mixture was stirred at room temperature for 2 hoursand quenched with water. After removal of all volatiles in vacuo, theresidue was extracted with 10% methanol in DCM twice. The combinedextracts were washed with brine, dried over anhydrous sodium sulfate.Concentration under vacuum gave a crude product, which was purified byreverse phase column to afford the title compound (1.08 g, yield: 63%).MS (ESI): 213 [M+H]⁺

c. 5-(2-Dimethylamino-ethyl-2-nitro-phenylamine

To a solution of [2-(3-fluoro-4-nitro-phenyl)-ethyl]-dimethyl-amine (800mg, 3.76 mmol) in MeOH (20 mL) was added ammonia hydroxide (5 mL). Thereaction mixture was heated at 100° C. under microwave heating for 1.5hours. The resulting yellow solid was collected and washed with water,dried under vacuum and combined with a previous batch to give the puretitle compound (560 mg, yield: 55%). MS (ESI): 210 [M+H]⁺.

Procedure 2H: Example—139

N-(2-((6-(3-(3,5-dimethoxyphenyl)-1methylureido)pyrimidin-4-ylamino)-5-(4-ethylpiperazin-1-yl)phenyl)acrylamide

a. 1-(6-chloropyrimidin-4-yl)-3-(3,5-dimethoxyphenyl)-1-methylurea

To a stirred solution of 6-chloro-N-methylpyrimidin-4-amine (1 g, 6.965mmol) in dioxane (10 mL) was added DIPEA (3.6 mL, 20.895 mmol) andtriphosgene (0.81 g, 2.786 mmol) under argon atmosphere at 0° C. Theresulting mixture was stirred for 1 h at 70° C., and then allowed tocool to room temperature. The resulting mixture was added via cannula toa solution of 3,5-dimethoxyaniline (1.2 g, 8.358 mmol) and DIPEA (1.2mL, 6.965 mmol) in dioxane (4 mL) under argon atmosphere at 0° C. Thereaction mixture was allowed to warm to room temperature and stirred for12 h. After completion of the reaction by TLC (Hexanes:EtOAc, 7:3),reaction mixture was diluted with ethyl acetate and a saturated aqueoussolution of NaHCO₃. The aqueous layer was separated and extracted withethyl acetate (2×50 mL). The combined organic phase washed with brine,dried on Na₂SO₄, filtered and concentrated under vacuum. The residue waspurified by silica gel column chromatography (Hexanes:EtOAc, 70:30) toafford 1-(6-chloropyrimidin-4-yl)-3-(3,5-dimethoxyphenyl)-1-methylurea(1.1 g, yield: 50%) as a white solid. ¹H NMR (CDCl₃, 300 MHz): δ 12.38(s, 1H), 8.71 (s, 1H), 6.99 (s, 1H), 6.79 (d, 2H), 6.26 (t, 1H), 3.81(s, 6H), 3.45 (s, 3H); MS (ESI): 323.10 [M+H]⁺.

b.1-(6-chloropyrimidin-4-yl)-3-(3,5-dimethoxyphenyl)-1-methyl-3-tert-butylcarbonate urea

NaH (0.124 g, 3.098 mmol) was added to a stirred solution of1-(6-chloropyrimidin-4-yl)-3-(3,5-dimethoxyphenyl)-1-methylurea (0.5 g,1.549 mmol) in anhydrous DMF (4 mL) under an argon atmosphere at 0° C.The resulting mixture was stirred for 15 min. and added theDi-tert-butyl dicarbonate (0.50 mL, 2.323 mmol) at 0° C. The resultingreaction mixture was allowed to warm to room temperature and stirred for16 h. The reaction mixture was diluted with ethyl acetate and an excessof cold water. The aqueous layer was separated and extracted with ethylacetate (3×25 mL). The combined organic phase was washed with brine,dried over Na₂SO₄, filtered, and concentrated under vacuum. The residuewas purified by silica gel column chromatography (Hexanes:EtOAc, 80:20)to afford the title compound (0.45 g, 69% of yield) as a white solid. ¹HNMR (CDCl₃, 300 MHz): δ 8.72 (d, 1H), 8.78 (d, 1H), 6.43-6.37 (m, 3H),3.77 (s, 6H), 3.49 (s, 3H), 1.40 (s, 9H); MS (ESI): 424.10 [M+2]⁺.

c. 1-(3,5-dimethoxyphenyl)-3-(6-((4-(4ethylpiperazin-1-yl)-2-nitrophenyl)amino)pyrimidin-4-yl)-3-methyl-1-tert-butylcarbonate urea

Pd₂(dba)₃ (0.095 g, 0.104 mmol) and Xantphos (0.1202 g, 0.208 mmol) wastaken in 10 mL of dry toluene in a seal tube under Argon atmosphere atroom temperature. The Argon gas purging was continued for additional5-10 min. Then1-(6-chloropyrimidin-4-yl)-3-(3,5-dimethoxyphenyl)-1-dimethyl-3-tert-butylcarbonate urea (0.43 g, 1.042 mmol) and4-(4-ethylpiperazin-1-yl)-2-nitroaniline (Procedure 2C, Steps a-c; 0.317g, 1.25 mmol) was added and the resulting reaction mixture was purgedwith argon gas for 5 min., and then Cs₂CO₃ (0.676 g, 2.08 mmol) wasadded. The argon gas purging was continued for additional 5 min. beforesealing the reaction vial. Then the reaction mixture was heated at 100°C. for 12 h. After completion of the reaction by TLC (DCM:MeOH, 98:2),reaction mass was partitioned between EtOAc and water. The aqueous layerwas extracted with EtOAc (2×25 mL) and the combined organic layer waswashed with water, brine, dried over Na₂SO₄ and evaporated under vacuum.The crude residue was purified by column chromatography on silica gel(DCM:MeOH/97:3) to afford the title compound (0.250 g, 37% of yield).¹HNMR (CDCl₃, 300 MHz): δ 9.30 (s, 1H), 8.58 (s, 1H), 8.17 (d, 1H), 7.59(d, 1H), 7.14 (s, 1H), 7.08 (dd, 1H), 6.42 (d, 2H), 6.36 (t, 1H), 3.75(s, 6H), 3.49 (s, 3H), 3.23 (t, 4H), 2.62 (t, 4H), 2.49 (q, 2H), 1.39(s, 9H), 1.14 (t, 3H); MS (ESI): 637.4 [M+H]⁺.

d. tert-butyl (6-(3-(3,5-dimethoxyphenyl)-1-methyl-3-tert-butylcarbonateureido)pyrimidin-4-yl)(4-(4-ethylpiperazin-1-yl)-2-nitrophenyl)carbamate

NaH (0.0314 g, 0.786 mmol) was added to a stirred solution of1-(3,5-dimethoxyphenyl)-3-(6-((4-(4-ethylpiperazin-1-yl)-2-nitrophenyl)amino)pyrimidin-4-yl)-3-methyl-1-tert-butylcarbonate urea (0.25 g, 0.393 mmol) in anhydrous DMF (4 mL) under anargon atmosphere at 0° C. The resulting mixture was stirred for 15 min.Then Di-tert-butyl dicarbonate (0.12 mL, 0.589 mmol) was added at 0° C.The resulting reaction mixture was allowed to warm to room temperatureand stirred for 18 h. The reaction mixture was diluted with ethylacetate and cold water. The aqueous layer was separated and extractedwith ethyl acetate (3×20 mL). The combined organic phase was washed withbrine, dried over Na₂SO₄, filtered and concentrated under vacuum. Theresidue was purified by silica gel column chromatography (DCM:MeOH,97:3) to afford the title compound (0.280 g, 96% of yield) as a brownsolid. ¹HNMR (CDCl₃, 300 MHz): δ 8.44 (s, 1H), 8.30 (s, 1H), 7.60 (d,1H), 7.14-7.03 (m, 2H), 6.48 (d, 1H), 6.44-6.21 (m, 2H), 3.75 (s, 6H),3.49 (s, 3H), 3.49 (t, 4H), 2.62 (t, 4H), 2.49 (q, 2H), 1.38 (d, 18H),1.14 (t, 3H); MS (ESI):737.5 [M+H]⁺.

e. tert-butyl (2-amino-4-(4-ethylpiperazin-1-yl)phenyl)(6-(3-(3,5-dimethoxyphenyl)-1-methyl-3-tert-butyl carbonateureido)pyrimidin-4-yl)carbamate

A mixture of tert-butyl(6-(3-(3,5-dimethoxyphenyl)-1-methyl-3-tert-butyl carbonateureido)pyrimidin-4-yl)(4-(4-ethylpiperazin-1-yl)-2-nitrophenyl)carbamate(0.280 g, 0.380 mmol) and Raney nickel (0.05 g) in mixture of MeOH andTHF (1:1) (10 mL) was stirred for 36 h at room temperature under ahydrogen atmosphere (balloon). The reaction mixture was filtered throughCelite pad. The filtrate was concentrated to afford the title compound(0.14 g, 52% of yield). MS (ESI): 707.7 [M+H]⁺.

f. tert-butyl(2-acrylamido-4-(4-ethylpiperazin-1-yl)phenyl)(6-(3-(3,5-dimethoxyphenyl)-1-methyl-3-tert-butylcarbonate ureido)pyrimidin-4-yl)carbamate

To a stirred solution of tert-butyl(2-amino-4-(4-ethylpiperazin-1-yl)phenyl)(6-(3-(3,5-dimethoxyphenyl)-1-methyl-3-tert-butylcarbonate ureido)pyrimidin-4-yl)carbamate (0.14 g, 0.183 mmol) inanhydrous DCM (5 mL) was added TEA (0.08 mL, 0.594 mmol) under argonatmosphere at 0° C. The resulting mixture was stirred for 15 min. andslowly added the acryloyl chloride (0.03 mL, 0.396 mmol) at 0° C. Theresulting reaction mixture was allowed to warm to room temperaturestirred for 3 h. The reaction mixture was diluted with DCM and water.The aqueous layer was separated and extracted with DCM (3×20 mL). Theorganic phase was washed with brine, dried over Na₂SO₄, filtered, andconcentrated. The residue was purified by silica gel columnchromatography (DCM:MeOH, 97:3) to afford the title compound (0.070 g,46% of yield) as a brown solid. MS (ESI): 761.4 [M+H]⁺.

g.N-(2-((6-(3-(3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-ethylpiperazin-1-yl)phenyl)acrylamide

TFA (0.35 mL, 5 vol) was slowly added to a stirred solution oftert-butyl(2-acrylamido-4-(4-ethylpiperazin-1-yl)phenyl)(6-(3-(3,5-dimethoxyphenyl)-1-methyl-3-tert-butylcarbonate ureido)pyrimidin-4-yl)carbamate (0.070 g, 0.124 mmol) in dryDCM (2 mL) under argon atmosphere at 0° C. The resulting reactionmixture was allowed to warm to room temperature and stirred for 24 h.Reaction progress was monitored by LCMS, after completion of thereaction, excess solvents were removed under reduced pressure. Theresulting residue was diluted with DCM and a saturated aqueous solutionof NaHCO₃. The aqueous layer was separated and extracted with DCM (3×10mL). The organic phase was washed with brine, dried on Na₂SO₄, filteredand concentrated under vacuum. The residue was purified by silica gelcolumn chromatography (DCM/MeOH, 97:3) to afford 45 mg of desiredproduct with HPLC purity 80%, which was purified by preparative HPLC(Conditions: Column: XBRIDGE-C18 (19.0×150 mm, 5 micron); (Mobile Phase:A; 0.1% TFA in Water, B; ACN) to afford the title compound (19 mg, 37%of yield) as a off-white solid. ¹HNMR (CDCl₃, 300 MHz): δ 12.85 (s, 1H),8.39 (s, 1H), 7.72 (s, 1H), 7.55 (s, 1H), 7.19 (d, 1H), 6.79 (s, 2H),6.75 (d, 1H), 6.64 (s, 1H), 6.39 (d, 1H), 6.22-6.13 (m, 2H), 5.80-5.73(m, 21H), 3.78 (s, 6H), 3.28 (t, 4H), 3.21 (s, 3H), 2.60 (t, 4H), 2.47(q, 2H), 1.12 (t, 3H); MS (ESI): 561.60 [M+H]⁺; HPLC: 96.04%, rt: 6.40min.

Example—140

N-(2-((6-(3-(2,6-dichlorophenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-ethylpiperazin-1-yl)phenyl)acrylamide

The compound was synthesized following the approach outlined inProcedure 2H (Example 139), substituting 2,6-dichloroaniline in step (a)and (2-(chloromethoxy)ethyl)trimethylsilane in step (b) to afford thetitle compound (18 mg, 3.2% of yield) as a off-white solid. ¹HNMR(CDCl₃, 300 MHz): δ 12.58 (s, 1), 8.39 (s, 1H), 7.75 (s, 1H), 7.61 (s,1H), 7.37 (d, 2H), 7.22 (d, 1H), 7.14 (t, 1H), 6.77 (dd, 1H), 6.67 (s,1H), 6.42 (d, 1H), 6.25-6.16 (m, 1H), 5.85 (s, 1H), 5.78 (d, 1H),3.35-3.24 (m, 7H), 2.62 (t, 4H), 2.49 (q, 2H), 1.14 (t, 3H); MS (ESI):569.10 [M]⁺; HPLC: 96.98%, rt: 3.49 min.

Example—141

N-(2-((6-(3-(2-chloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-ethylpiperazin-1-yl)phenyl)acrylamide

The compound was synthesized following the approach outlined inProcedure 2H (Example 139), substituting 2-chloro-3,5-dimethoxyaniline(procedure shown below) in step (a) to afford the title compound (20 mg,6.8% of yield) as a off-white solid. 1H-NMR (CDCl₃, 400 MHz): δ 13.46(s, 1H), 8.43 (s, 1H), 7.79-7.71 (m, 2H), 7.56 (s, 1H), 7.18 (d, 1H),6.74 (dd, 1H), 6.61 (s, 1H), 6.38 (d, 1H), 6.26 (d, 1H), 6.21-6.15 (m,1H), 5.79-5.74 (m, 2H), 3.86 (s, 3H), 3.81 (s, 3H), 3.28 (t, 4H), 3.23(s, 3H), 2.59 (t, 4H), 2.47 (q, 2H), 1.12 (t, 3H); MS (ESI): 595.15[M]⁺; HPLC: 98.14%, rt: 3.49 min.

Preparation of 2-chloro-3,5-dimethoxyaniline

a. N-(3,5-dimethoxyphenyl)acetamide

Acetic anhydride (6.5 mL) was added slowly to a stirred solution of3,5-dimethoxy aniline (10 g, 65.359 mmol) in toluene (50 mL) under argonatmosphere at room temperature and resulting reaction mixture wasstirred for 15 h. After completion of the reaction, reaction was dilutedwith Hexane and resulted precipitate was collected by filtration anddried under vacuum to afford the title compound (12.5 g, 98% of yield)as a off-white solid. ¹HNMR (CDCl₃, 300 MHz): δ 7.38 (s, 1H), 6.75 (d,2H), 6.23 (s, 1H), 3.76 (s, 6H), 2.15 (s, 3H); MS (ESI):196. [M+H]⁺.

b. N-(2-chloro-3,5-dimethoxyphenyl)acetamide

To a stirred solution of N-(3,5-dimethoxy-phenyl)-acetamide (5 g, 25.64mmol) in acetic acid (17 mL) was added 32% aqueous hydrochloric acidsolution (14 mL), followed by a solution of sodium chlorate (1.16 g, 11mmol) in water (1.5 mL) at 0° C. The resulting reaction mixture wasstirred for 30 min. at 0° C. Thereafter reaction mixture was poured intoice water and made it basic with K₂CO₃ powder. The precipitate wasfiltered off and washed with water. The residue was purified by silicagel column chromatography (Hexane/EtOAc, 88:12) to afford the titlecompound (1.8 g, 31% of yield) as a white solid. ¹HNMR (DMSO-d₆, 300MHz): δ 9.36 (s, 1H), 7.03 (d, 1H), 6.53 (d, 1H), 3.84 (s, 3H), 3.74 (s,3H), 2.08 (s, 3H); MS (ESI): 230.2 [M+H]⁺.

c. 2-chloro-3,5-dimethoxyaniline

Potassium hydroxide (2.19 g, 39.18 mmol) was added to a solution ofN-(2-chloro-3,5-dimethoxyphenyl)acetamide (1.8 g, 7.837 mmol) in EtOH(100 mL) and water (10 mL) and the reaction mixture heated to reflux for12 h. Excess EtOH was removed under reduced pressure to obtain aresidue. The residue was then partitioned between water and diethylether. The organic layer was separated, dried over sodium sulfate,filtered and concentrated under vacuum to afford the title compound (1.2g, 82% of yield) as a white solid. ¹HNMR (CDCl₃, 300 MHz): δ 5.97 (s,2H), 4.08 (brs, 2H), 3.84 (s, 3H), 3.75 (s, 3H); MS (ESI):188.1 [M+H]⁺.

Procedure 21: Example—142

N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(2-methoxyethoxy)phenyl)acrylamide

a. 4-(2-methoxyethoxy)-2-nitroaniline

Powdered and dried Potassium carbonate (3.58 g, 0.025 mol) was added toa solution of 4-amino-3-nitrophenol (2 g, 0.012 mol) in DMF (20 mL) at0° C. under nitrogen atmosphere. To this, 1-bromo-2-methoxyethane (1.34mL, 0.014 mol) was added dropwise and the resultant reaction mixture wasrefluxed for overnight. Then, the reaction mixture was filtered throughcelite. The filterate was concentrated and diluted with ethyl acetateand water. The aqueous layer was separated and extracted with ethylacetate (3×30 mL). The organic phase was washed with brine, dried onNa₂SO₄, filtered and concentrated under vacuum. The residue was purifiedby silica gel column chromatography to obtain the title compound (0.6 g,yield: 24%) as a solid. MS (ESI): 213.15 [M+H]⁺.

b.N⁴-(4-(2-methoxyethoxy)-2-nitrophenyl)-N⁶-methylpyrimidine-4,6-diamine

4-(2-methoxyethoxy)-2-nitroaniline (0.6 g, 2.830 mmol) and6-chloro-N-methylpyrimidin-4-amine (0.404 g, 2.830 mmol) were taken in10 mL of dry toluene in a seal tube under Argon atmosphere at roomtemperature. The Argon gas purging was continued for additional 5-10min. Then Cs₂CO₃ (2.3 g, 7.075 mmol, 2.5 eq) and Xantphos (0.490 g,0.849 mmol) were added and the resulting reaction mixture was purgedwith argon gas for 5 min, followed by Pd₂(dba)₃ (0.518 g, 0.566 mmol)was added. The argon gas purging was continued for additional 5 minbefore sealing the reaction vial. Then the reaction mixture was heatedat 100° C. for 7 h. After completion of the reaction by TLC (DCM:MeOH,98:2), reaction mass was filtered through celite and the filtrate wasevaporated under vacuum to get a crude residue. The crude residue waspurified by column chromatography on silica gel to afford the titlecompound (0.93 g, yield: 70%); MS (ESI): 320.3 [M+H]⁺.

c.3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-(6-((4-(2-methoxyethoxy)-2-nitrophenyl)amino)pyrimidin-4-yl)-1-methylurea

To a stirred solution of 2,6-dichloro-3,5-dimethoxyaniline (500 mg,2.252 mmol) in dioxane (10 mL) was added 20% phosgene in toluene (4.4mL, 9.0 mmol) under argon atmosphere at 0° C. The resulting mixture wasstirred for 6 h at 90° C., and then allowed to cool to room temperature.The solvents were removed and the residue was dissolved in toluene (10mL). To this, was addedN4-(4-(2-methoxyethoxy)-2-nitrophenyl)-N6-methylpyrimidine-4,6-diamine(0.718 g, 8.358 mmol, 1.0 eq). The resultant reaction mixture was thenrefluxed for 6 h. After completion of the reaction by TLC(Hexanes:EtOAc, 7:3), reaction mixture was cooled to room temperature,concentrated under vacuum to obtain a crude reaction mixture. The solidprecipitated on addition of ethyl acetate to the crude reaction mixturewas filtered, washed with ether and pentane to afford the title compound(0.285, yield: 32%) as a white solid. MS (ESI): 567.0 M+H.

d. tert-butyl(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-tert-butyl carbonateureido)pyrimidin-4-yl)(4-(2-methoxyethoxy)-2-nitrophenyl)carbamate

DMAP (0.025 g, 0.2 mmol), and Di-tert-butyl dicarbonate (0.438 g, 2.009mmol) was added to a stirred solution of3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-(6-((4-(2-methoxyethoxy)-2-nitrophenyl)amino)pyrimidin-4-yl)-1-methylurea (0.285 g, 0.502 mmol) in anhydrous THF (10mL) under an argon atmosphere at 0° C. The resulting mixture wasrefluxed for 3-4 h. After completion of the reaction by TLC(Hexanes:EtOAc, 1:1), reaction mixture was cooled to room temperature,concentrated under vacuum to obtain a crude residue. The residue waspurified by silica gel column chromatography to afford the titlecompound (0.35 g, yield: 88%) as an off-white solid. MS (ESI): 767.1[M+H]⁺.

e. ter-butyl(2-amino-4-(2-methoxyethoxy)phenyl)(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-tert-butylcarbonate ureido)pyrimidin-4-yl)carbamate

Raney nickel (0.05 g) was added to a solution of tert-butyl(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-tert-butyl carbonateureido)pyrimidin-4-yl)(4-(2-methoxyethoxy)-2-nitrophenyl)carbamate(0.350 g, 0.456 mmol) in mixture of THF and MeOH (10 mL) and theresultant reaction mixture was stirred for 14 h at room temperatureunder a hydrogen atmosphere (balloon). The reaction mixture was filteredthrough Celite pad. The filtrate was concentrated to afford a cruderesidue. The residue was purified by silica gel column chromatography(MeOH:DCM, 5:95) to afford the title compound (0.26 g, yield: 77%) assolid. MS (ESI): 737.2 [M+H]⁺.

f. tert-butyl(2-acrylamido-4-(2-methoxyethoxy)phenyl)(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-tert-butylcarbonate ureido)pyrimidin-4-yl)carbamate

To a stirred solution of tert-butyl(2-amino-4-(2-methoxyethoxy)phenyl)(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-tert-butylcarbonate ureido)pyrimidin-4-yl)carbamate (0.26 g, 0.352 mmol) inanhydrous DCM (6 mL) was added TEA (0.09 mL, 0.704 mmol) under argonatmosphere at 0° C. The resulting mixture was stirred for 15 min. andslowly added the acryloyl chloride (0.04 mL, 0.528 mmol) at 0° C. Theresulting reaction mixture was allowed to warm to room temperature andstirred for 2 h. The reaction mixture was diluted with DCM and water.The aqueous layer was separated and extracted with DCM (3×30 mL). Theorganic phase was washed with brine, dried over Na₂SO₄, filtered, andconcentrated. The residue was purified by silica gel columnchromatography to afford the title compound (0.200 g, yield: 72%) as asolid. MS (ESI): 791.2 [M+H]⁺.

g.N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(2-methoxyethoxy)phenyl)acrylamide

TFA (0.38 mL, 5.05 mmol) was slowly added to a stirred solution oftert-butyl(2-acrylamido-4-(2-methoxyethoxy)phenyl)(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-tert-butylcarbonate ureido)pyrimidin-4-yl)carbamate (0.2 g, 0.252 mmol) in dry DCM(2 mL) under argon atmosphere at 0° C. The resulting reaction mixturewas allowed to warm to room temperature and stirred for 12 h. Reactionprogress was monitored by LCMS. After completion of the reaction, excesssolvents were removed under reduced pressure. The crude solid was washedwith ether to afford the title compound (86 mg, yield: 58%) as a whitesolid. ¹HNMR (DMSO-d6, 400 MHz): δ12.01 (s, 1H), 9.60 (s, 1H), 8.85 (s,1H), 8.33 (s, 1H); 7.42 (s, 1H), 7.34 (d, 1H), 6.89 (s, 1H), 6.79 (d,1H), 6.51 (dd, 1H), 6.23 (d, 2H), 5.72 (d, 1H), 4.08 (m, 2H), 3.93 (s,6H), 3.66 (m, 2H), 3.31 (s, 3H), 3.23 (s, 3H). MS (ESI): 591.3 [M+H]⁺;HPLC: 96.04%, rt: 3.72 min.

Example: 144

N-(2-((6-(3-(2-chloro-6-fluoro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-ethylpiperazin-1-yl)phenyl)acrylamide

The title compound was synthesized following the approach outlined inProcedure 21 (Example 142), substitutingN⁴-(4-(4-ethylpiperazin-1-yl)-2-nitrophenyl)-N⁶-methylpyrimidine-4,6-diamine(Procedure 2C, Example 108, Steps a-d) and2-chloro-6-fluoro-3,5-dimethoxyaniline (Procedure shown below) in step(c) to afford the pen-ultimate title compound (0.46 mg, yield: 2.6% overfive steps) as an off-white solid. MS (ESI): 813.1 [M+H]⁺. The freeamine (46 mg, 0.075 mmol) was dissolved in ethyl acetate: DCM: MeOHmixture and treated with phosphoric acid (7 mg, 0.075 mmol). Afterstirring for 1 h, the solid precipitated was filtered and washed withether and pentane to afford the title compound (34.7 mg, yield: 65%) asan off-white solid. ¹HNMR (DMSO-d6, 300 MHz): δ 12.15 (s, 1H), 9.60 (s,1H), 8.76 (s, 1H), 8.32 (s, 1H), 7.29 (d, 2H), 6.9 (d, 2H), 6.81 (dd,1H), 6.51-6.45 (m, 1H), 6.25-6.20 (m, 2H), 5.72 (d, 1H), 3.91 (s, 3H),3.89 (s, 3H), 3.23 (t, 4H), 3.14 (t, 4H), 2.54-2.50 (m, 5H), 1.05 (t,3H); MS (ESI): 613.2 (M−H₃PO₄]⁺; HPLC: 98.6%, rt: 6.13 min.

Preparation of 2-chloro-6-fluoro-3,5-dimethoxyaniline

a. methyl 2-fluoro-3,5-dimethoxybenzoate

A suspension of Selectfluor (48.9 g, 0.15 mol) in acetonitrile (1.1 L)was added to a solution of methyl-3,5-dimethoxy benzoate (20 g, 0.10mol) in acetonitrile at 0° C. under nitrogen atmosphere. The resultingreaction mixture was warmed to room temperature and stirred overnight.The reaction mixture was concentrated under vacuum, diluted withsaturated sodium carbonate solution and ethyl acetate. The aqueous layerwas separated and extracted with ethyl acetate (3×200 mL). The organicphase was washed with brine, dried over Na₂SO₄, filtered, andconcentrated. The residue was purified by silica gel columnchromatography (gradient hexane/ether 30:1 to 4:1) to afford the titlecompound (4 g, yield: 16.9%).

b. methyl 2-chloro-6-fluoro-3,5-dimethoxybenzoate

SO₂C₂ (2.20 g, 0.016 mol) was added dropwise to a solution ofmethyl-2-fluoro-3,5-dimethoxy benzoate (3.5 g, 0.016 mol) inacetonitrile (40 mL) at 0° C. under nitrogen atmosphere. The resultingreaction mixture was warmed to room temperature slowly and stirred for 1h. The reaction mixture was quenched with saturated sodium bicarbonatesolution, and extracted with ethyl acetate (3×30 mL). The organic phasewas washed with brine, dried over Na₂SO₄, filtered, and concentrated.The residue was purified by silica gel column chromatography usingeluent (gradient hexane/ether (20:1) to hexane/ether (5:1) to afford thetitle compound (2.7 g, yield: 67%) as a solid.

c. 2-chloro-6-fluoro-3,5-dimethoxybenzoic acid

A suspension of methyl 2-chloro-6-fluoro-3,5-dimethoxybenzoate (2.7 g,0.010 mol) and sodium hydroxide (1.088 g, 0.0272) in anhydrous ethanol(30 mL) was refluxed for 24 h. The resulting reaction mixture was cooledto room temperature and concentrated under vacuum to get a cruderesidue. The crude residue was dissolved in water and extracted withether (3×30 mL). The aqueous layer was acidified with conc.HCl and theprecipitated solid was filtered, washed with cold water and dried invacuo to afford the title compound (1.8 g, yield: 71%) as a solid.

d. 2-chloro-6-fluoro-3,5-dimethoxyaniline

A suspension of 2-chloro-6-fluoro-3,5-dimethoxybenzoic acid (1.8 g,0.0077 mol) and triethyl amine (0.934 g, 0.0092 mol) in tert-BuOH (50mL) was stirred for 5 min. To the resulting reaction mixture, Diphenylphosphoryl azide (2.53 g, 0.0092 mol) was added and heated up to 82° C.and kept at this temperature for overnight. The reaction mixture wasthen concentrated in vacuo to obtain a crude residue. The crude residuewas dissolved in dichloromethane (20 mL) and cooled to 0° C. TFA (4 mL)was added to the reaction mixture and the resultant reaction mixture wasthen stirred at room temperature for 2 h. The solvents were removedunder vacuum and the crude residue was diluted with ethyl acetate andsaturated sodium carbonate solution. The aqueous layer was separated andextracted with ethyl acetate (3×30 mL). The organic phase was washedwith brine, dried over Na₂SO₄, filtered, and concentrated. The residuewas purified by silica gel column chromatography using the eluent(gradient hexane to hexane-ether (65:35)) to afford the title compound(0.95 g, yield: 60%) as solid. MS (ESI): 205.7 [M+H]⁺.

Procedure 2J—Example 145

N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(hydroxymethyl)phenyl)acrylamide

a.1-(6-chloropyrimidin-4-yl)-3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-tert-butylcaronate urea

DMAP (0.080 g, 0.655 mmol) and Di-tert-butyl dicarbonate (2.9 mL, 12.6mmol) was added to a stirred solution of1-(6-chloropyrimidin-4-yl)-3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylurea(Procedure 2E, step b; 2.6 g, 6.632 mmol) in anhydrous THF (20 mL) underan argon atmosphere at 0° C. The resulting mixture was then refluxed for2 h. After completion of the reaction by TLC (EtOAc:Hexane 3:7),reaction mixture was cooled to room temperature, concentrated undervacuum to obtain a crude residue. The residue was purified by silica gelcolumn chromatography to afford the title compound (2.5 g, yield: 69.4%)as a solid. ¹H-NMR (CDCl₃, 300 MHz): δ 8.73 (d, 1H), 7.95 (d, 1H), 6.61(s, 1H), 3.95 (s, 6H), 3.63 (s, 3H), 1.35 (s, 9H).

b. methyl4-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-tert-butylcarbonate ureido)pyrimidin-4-yl)amino)-3-nitrobenzoate

Methyl 4-amino-3-nitrobenzoate (0.956 g, 0.004 mol) and1-(6-chloropyrimidin-4-yl)-3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-tert-butylcaronate urea (2 g, 0.004 mol) were taken in 10 mL of dry toluene in aseal tube at room temperature and Argon gas was purged for 5-10 min.Then Cs₂CO₃ (3.25 g, 0.01 mol) and Xantphos (0.46 g, 0.0008 mol) wereadded and the resulting reaction mixture was purged with argon gas for 5min, followed by Pd₂(dba)₃ (0.36 g, 0.0004 mol) was added. The argon gaspurging was continued for additional 5 min before sealing the reactionvial. Then the reaction mixture was heated at 100° C. for 12 h. Aftercompletion of the reaction by TLC, reaction mass was filtered throughcelite and the filterate was evaporated under vacuum to get a cruderesidue. The crude residue was purified by column chromatography onsilica gel to afford the title compound (2.10 g, yield: 79%); ¹H-NMR(CDCl₃, 300 MHz): δ 10.3 (s, 1H), 9.00-8.95 (m, 2H), 8.75 (s, 1H), 8.22(dd, 1H), 7.63 (s, 1H), 6.61 (s, 1H), 3.95 (s, 6H), 3.94 (s, 3H), 3.67(s, 3H), 1.39 (s, 9H).

c. methyl 4-((tert-butoxycarbonyl)(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-tert-butylureido)pyrimidin-4-yl)amino)-3-nitrobenzoate

DMAP (0.039 g, 0.322 mmol) and Di-tert-butyl dicarbonate (1.48 mL, 6.45mmol) was added to a stirred solution of methyl4-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-tert-butylcarbonate ureido)pyrimidin-4-yl)amino)-3-nitrobenzoate (2.1 g, 3.22mmol) in anhydrous THF (5 mL) under an argon atmosphere at 0° C. Theresulting mixture was refluxed for 12 h. After completion of thereaction by TLC (EtOAc:Hexane 40:60), reaction mixture was cooled toroom temperature, concentrated under vacuum to obtain a crude residue.The residue was purified by silica gel column chromatography to affordthe title compound (2.1 g, yield: 87%) as a solid. MS (ESI): 751.0 [M]⁺.

d. tert-buty(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-tert-butyl carbonateureido)pyrimidin-4-yl) (4-(hydroxymethyl)-2-nitrophenyl)carbamate

Lithium borohydride (0.049 g, 2.26 mmol) was added to a solution ofmethyl4-((tert-butoxycarbonyl)(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-tert-butylureido)pyrimidin-4-yl)amino)-3-nitrobenzoate (1.7 g, 2.26 mmol) inanhydrous THF (18 mL) under argon atmosphere at 0° C. The resultingreaction mixture was then allowed to warm to room temperature by 2.5 h.The reaction mixture was then quenched with ice-water and diluted withethylacetate. The aqueous layer was separated and extracted with ethylacetate (3×20 mL). The organic phase was washed with brine, dried overNa₂SO₄, filtered and concentrated under vacuum. The residue was purifiedby silica gel column chromatography to afford the title compound (0.480g, yield: 29%) as a solid. MS (ESI): 723.2 [M]⁺.

e. tert-butyl(4-(((tert-butyldimethylsilyl)oxy)methyl)-2-nitrophenyl)(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-tert-butylcarbonate ureido)pyrimidin-4-yl)carbamate

Imidazole (0.071 g, 1.051 mmol) and TBDMS-Cl (0.118 g, 0.787 mmol) wereadded to a stirred solution of tert-butyl(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-tert-butyl carbonateureido)pyrimidin-4-yl)(4-(hydroxymethyl)-2-nitrophenyl)carbamate (0.380g, 0.525 mmol) in anhydrous dichloromethane (5 mL) under an argonatmosphere at 0° C. The resultant reaction mixture was stirred for 4 hat room temperature. The reaction mixture was then diluted with DCM andwater. The aqueous layer was separated and extracted with DCM (3×10 mL).The organic phase was washed with brine, dried over Na₂SO₄, filtered andconcentrated under vacuum. The residue was purified by silica gel columnchromatography to afford the title compound (0.330 g, yield: 75%) as asolid. MS (ESI): 837.4 [M]⁺.

f. tert-butyl(2-amino-4-((tert-butyldimethylsilyl)oxy)methyl)phenyl)(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-tert-butylcarbonate ureido)pyrimidin-4-yl)carbamate

Raney nickel (0.06 g) was added to a solution of tert-butyl(4-(((tert-butyldimethylsilyl)oxy)methyl)-2-nitrophenyl)(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-tert-butylcarbonate ureido)pyrimidin-4-yl)carbamate (0.3 g, 0.358 mmol) in MeOH (5mL) and the resultant reaction mixture was stirred for 12 h at roomtemperature under a hydrogen atmosphere (balloon). The reaction mixturewas filtered through Celite pad. The filtrate was concentrated to afforda crude residue which was then purified by column chromatography toafford the title compound (0.180 g, yield: 64%) as a solid. MS (ESI):807.2 [M]⁺.

g. tert-butyl(2-acrylamido-4-(((tert-butyldimethylsilyl)oxy)methylphenyl)(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-tert-butylcarbonate ureido)pyrimidin-4-yl)carbamate

To a stirred solution of tert-butyl(2-amino-4-(((tert-butyldimethylsilyl)oxy)methyl)phenyl)(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-tert-butylcarbonate ureido)pyrimidin-4-yl)carbamate (0.180 g, 0.223 mmol) inanhydrous DCM (5 mL) was added TEA (0.08 mL, 0.557 mmol) under argonatmosphere at 0° C. The resulting mixture was stirred for 15 min. andslowly added the acryloyl chloride (0.03 mL, 0.334 mmol) at 0° C. Theresulting reaction mixture was allowed to warm to room temperature andstirred for 4 h. The reaction mixture was then concentrated and theresidue was purified by silica gel column chromatography to afforddesired the title compound (0.130 g, yield: 68%) as a solid. MS (ESI):861.3 [M]f.

h.N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(hydroxymethyl)phenyl)acrylamide

TFA (1.0 mL) was slowly added to a stirred solution of tert-butyl(2-acrylamido-4-(((tert-butyldimethylsilyl)oxy)methyl)phenyl)(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-tert-butylcarbonate ureido)pyrimidin-4-yl)carbamate (0.130 g, 0.150 mmol) in dryDCM (3 mL) under argon atmosphere at 0° C. The resulting reactionmixture was allowed to warm to room temperature and stirred for 13 h.Reaction progress was monitored by LCMS, after completion of thereaction, excess solvents were removed under reduced pressure. Theresulting residue was diluted with DCM and a saturated aqueous solutionof NaHCO₃. The aqueous layer was separated and extracted with DCM (3×10mL). The organic phase was washed with brine, dried over Na₂SO₄,filtered and concentrated under vacuum. The residue was purified bysilica gel column chromatography to afford 95 mg of desired product withHPLC purity 84%, which was then purified by preparative HPLC(Conditions: Column: X bridge C18 (19 mm×150 mm, 5 μm); (Mobile Phase:A; 0.01% TFA in Water, B; ACN) to afford the title compound (16 mg,yield: 16%) as a white solid. ¹H-NMR (DMSO-d6, 400 MHz): δ 12.08 (s,1H), 9.70 (s, 1H), 8.95 (s, 1H), 8.37 (s, 1H), 7.64 (s, 1H), 7.49 (d,1H), 7.15 (s, 1H), 6.90 (s, 1H), 6.53-7.46 (q, 1H), 6.32 (s, 1H), 6.25(d, 1H), 5.74 (d, 1H), 5.42 (s, 1H), 5.35 (s, 1H), 4.49 (s, 2H), 3.94(s, 6H), 3.25 (s, 3H); MS (ESI): 547.0 [M+H]*; HPLC: 97.4%, rt: 3.83min.

Example 147

N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(methoxymethyl)phenyl)acrylamide

The title compound was synthesized following the approach outlined inProcedure 21 (Example 142), substitutingN⁴-(4-(methoxymethyl)-2-nitrophenyl)-N⁶-methylpyrimidine-4,6-diamine(procedure shown below) in step (c) to afford the title compound (5.0mg, yield: 1.8%) as an off-white solid. 1H-NMR (CDCl₃, 400 MHz): δ 12.30(s, 1H), 8.41 (s, 1H), 7.86 (s, 1H), 7.81 (s, 1H), 7.52 (d, 1H), 7.49(s, 1H), 7.30 (d, 1H), 6.54 (s, 1H), 6.46 (d, 1H), 6.26 (dd, 1H), 6.07(s, 1H), 5.83 (d, 1H), 5.18 (s, 2H), 3.93 (s, 6H), 3.82 (s, 3H), 3.34(s, 3H); MS (ESI): 561.1 [M+H]⁺; HPLC: 95.76%, rt: 4.36 min.

Preparation of(4-(4-(methoxymethyl)-2-nitrophenyl)-N⁶-methylpyrimidine-4,6-diamine

a. methyl 4-amino-3-nitrobenzoate

Thionyl chloride (19.4 g, 164.85 mmol) was added to a solution of4-amino-3-nitrobenzoic acid (20 g, 109.89 mmol) in methanol (200 mL) at0° C. The resulting mixture was then refluxed for 12 h. The reactionmixture was allowed to cool to room temperature. The yellow solidprecipitated was filtered and dried to afford the title compound (22 g,yield: 100%) as a solid. 1H-NMR (CDCl₃, 300 MHz): δ 8.85 (d, 1H), 7.99(dd, 1H), 6.83 (d, 1H), 6.40 (s, 2H), 3.90 (s, 3H).

b. methyl 4-((6-(methylamino)pyrimidin-4-yl)amino)-3-nitrobenzoate

6-chloro-N-methylpyrimidin-4-amine, 3 (3 g, 20.97 mmol) and methyl4-amino-3-nitrobenzoate (3.9 g, 20.97 mmol) were taken in toluene (5 mL)in a seal tube under Argon atmosphere at room temperature. The Argon gaswas purged for 5-10 min. Then Cs₂CO₃ (17.0 g, 52.4 mmol) and Xantphos(3.6 g, 6.29 mmol) were added and the resulting reaction mixture waspurged with argon gas for 5 min, followed by Pd₂(dba)₃ (3.8 g, 4.19mmol) was added. The argon gas purging was continued for additional 5min before sealing the reaction vial. Then the reaction mixture washeated at 110° C. for 16 h. After completion of the reaction by TLC, thereaction mixture was cooled to rt, filtered through celite bed and thefiltrate was evaporated under vacuum to get a crude residue. The cruderesidue was purified by column chromatography on silica gel to affordthe title compound (3.2 g, yield: 51%) as a solid; MS (ESI): 304.2[M+H]⁺.

c. methyl 4-((tert-butoxycarbonyl)(6-((tert-butoxycarbonyl)(methyl)amino)pyrimidin-4-yl)amino)-3-nitrobenzoate

DMAP (0.497 g, 4.078 mmol) and Di-tert-butyl dicarbonate (8.89 g, 40.78mmol) were added to a stirred solution of intermediate-4 (3.1 g, 10.197mmol) in anhydrous THF (35 mL) under an argon atmosphere at roomtemperature. The resulting mixture was refluxed for 2-3 h. Aftercompletion of the reaction by TLC, reaction mixture was cooled to roomtemperature, concentrated under vacuum to obtain a crude residue. Theresidue was purified by silica gel column chromatography to afford thetitle compound (1.9 g, yield: 37%) as a solid. MS (ESI): 504.0 [M+H]⁺.

d. tert-butyl (6-((tert-butoxycarbonyl)(4-(hydroxymethyl)-2-nitrophenyl)amino)pyrimidin-4-yl)(methyl) carbamate

Lithium borohydride (0.157 g, 7.14 mmol) was added to a solution ofmethyl4-((tert-butoxycarbonyl)(6-((tert-butoxycarbonyl)(methyl)amino)pyrimidin-4-yl)amino)-3-nitrobenzoate(1.8 g, 3.57 mmol) in THF (20 mL) under an argon atmosphere at 0° C. Theresulting mixture was then allowed to warm to rt and stirred for 12 h.The reaction mixture was then quenched with ice-water and diluted withethyl acetate. The aqueous layer was separated and extracted with ethylacetate (3×40 mL). The organic phase was washed with brine, dried overNa₂SO₄, filtered, and concentrated to get a crude residue. The residuewas purified by silica gel column chromatography to afford desiredintermediate-6 (1.2 g, 75%) as a solid. MS (ESI): 476.1 [M+H]⁺.

e. tert-butyl (6-((tert-butoxycarbonyl)(4-(methoxymethyl)-2-nitrophenyl)amino)pyrimidin-4-yl)(methyl)carbamate

To a solution of tert-butyl(6-((tert-butoxycarbonyl)(4-(hydroxymethyl)-2-nitrophenyl)amino)pyrimidin-4-yl)(methyl)carbamate (15 g, 3.488 mmol) in acetone (20 mL)under an argon atmosphere at 0° C., was added potassium carbonate (0.48g, 6.97 mmol) followed by dimethyl sulfate (0.87 g, 6.97 mmol). Theresulting reaction mixture was then refluxed for 24 h and then allowedto cool to rt. The reaction mixture was concentrated under vacuum toobtain a crude residue. The residue was purified by silica gel columnchromatography to afford the title compound (0.3 g, yield: 19%) as asolid. MS (ESI): 490.55 [M+H]⁺.

f. N⁴-(4-(methoxymethyl)-2-nitrophenyl)-N⁶-methylpyrimidine-4,6-diamine

TFA (5 mL) was slowly added to a stirred solution of tert-butyl(6-((tert-butoxycarbonyl)(4-(methoxymethyl)-2-nitrophenyl)amino)pyrimidin-4-yl)(methyl)carbamate(0.85 g) in dry DCM (10 mL) under argon atmosphere at 0° C. Theresulting reaction mixture was allowed to warm to room temperature andstirred for 1 h. After completion of the reaction, excess solvents wereremoved under reduced pressure and washed with ether to afford the titlecompound (0.7 g crude) as a solid. MS (ESI): 290.2 [M+H]⁺.

Procedure 2K—Example 148

N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-methylpiperazine-1-carbonyl)phenyl)acrylamide

a.4-((tert-butoxycarbonyl)(6-(3-(tert-butoxycarbonyl)-3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-3-nitrobenzoicacid

Lithium hydroxide (0.08 g, 2.99 mmol) was added to a solution of methyl4-((tert-butoxycarbonyl)(6-(3-(tert-butoxycarbonyl)-3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-3-nitrobenzoate(1.5 g, 1.997 mmol) (Procedure 2J, Step c) in a mixture of THF (10 mL)and water (2 mL) at 0° C. The resulting reaction mixture was allowed towarm to room temperature and stirred for 5 h. After completion of thereaction by TLC (EtOAc:Hexane 3:7), the reaction mixture wasconcentrated under vacuum to get a crude residue. The crude residue wasdissolved in water and extracted with ether (3×30 mL). The aqueous layerwas acidified with 10% citric acid solution and the precipitated solidwas filtered, washed with cold water and dried in vacuo to afford thetitle compound (1.3 g, yield: 93%) as a light brown solid. MS (ESI):737.2 [M+H]⁺.

b. tert-butyl (2-nitro-4-(4-methylpiperazine-1-carbonyl)phenyl)(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-3-tert-butoxycarbonyl-1-methylureido)pyrimidin-4-yl)carbamate

DIPEA (0.3 mL, 1.62 mmol), HATU (0.515 g, 1.355 mmol) and N-methylpiperazine, 6 (0.09 mL, 0.813 mmol) were added to a solution of4-((tert-butoxycarbonyl)(6-(3-(tert-butoxycarbonyl)-3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-3-nitrobenzoicacid (0.4 g, 0.542 mmol) in DMF (5 mL) at 0° C. under argon atmosphere.The resulting reaction mixture was then allowed to warm to rt andstirred for 18 h. After completion of the reaction by TLC (MeOH:DCM1:19), water was added to the reaction mixture. The precipitated crudesolid was filtered, dried and purified by silica gel columnchromatography to obtain the title compound (0.27 g, yield: 61%) as asolid. MS (ESI): 819.1 [M+H]⁺.

c. tert-butyl (2-amino-4-(4-methylpiperazine-1-carbonyl)phenyl)(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-3-tert-butoxycarbonyl-1-methylureido)pyrimidin-4-yl)carbamate

Raney nickel (0.06 g) was added to a solution of tert-butyl(2-nitro-4-(4-methylpiperazine-1-carbonyl)phenyl)(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-3-tert-butoxycarbonyl-1-methylureido)pyrimidin-4-yl)carbamate(0.27 g, 0.329 mmol) in a mixture of MeOH (4 mL) and THF (4 mL) and theresultant reaction mixture was stirred for 20 h at room temperatureunder a hydrogen atmosphere. The reaction mixture was filtered throughCelite bed. The filtrate was concentrated, to afford crude titlecompound (0.22 g, yield: 84%) as a solid. MS (ESI): 789.4 [M+H]⁺.

d. tert-butyl (2-acrylamido-4-(4-methylpiperazine-1-carbonyl)phenyl)(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-3-tert-butoxycarbonyl-1-methylureido)pyrimidin-4-yl)carbamate

To a stirred solution of tert-butyl(2-amino-4-(4-methylpiperazine-1-carbonyl)phenyl)(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-3-tert-butoxycarbonyl-1-methylureido)pyrimidin-4-yl)carbamate(0.22 g, 0.278 mmol) in anhydrous DCM (5 mL) was added TEA (0.08 mL,0.557 mmol) under argon atmosphere at 0° C. The resulting mixture wasstirred for 15 min. and slowly added the acryloyl chloride (0.037 g,0.417 mmol) at 0° C. The resulting reaction mixture was allowed to warmto room temperature and stirred for 2 h. The reaction mixture wasquenched with a saturated sodium bicarbonate solution and diluted withDCM. The aqueous layer was separated and extracted with DCM (3×20 mL).The organic phase was washed with brine, dried over Na₂SO₄, filtered,and concentrated to get a crude residue. The residue was purified bysilica gel column chromatography to afford the title compound (0.060 g,yield: 25%) as a solid. MS (ESI): 843.3 [M+H]⁺.

e.N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-methylpiperazine-1-carbonyl)phenyl)acrylamide

TFA (0.2 mL) was slowly added to a stirred solution of tert-butyl(2-acrylamido-4-(4-methylpiperazine-1-carbonyl)phenyl)(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-3-tert-butylcarbonyl-1-methylureido)pyrimidin-4-yl)carbamate(0.060 g, 0.071 mmol) in dry DCM (2 mL) under argon atmosphere at 0° C.The resulting reaction mixture was allowed to warm to room temperatureand stirred for 18 h. Reaction progress was monitored by LCMS, aftercompletion of the reaction, excess solvents were removed under reducedpressure. The resulting residue was diluted with DCM and quenched with asaturated aqueous solution of NaHCO₃. The aqueous layer was separatedand extracted with DCM (3×10 mL). The organic phase was washed withbrine, dried over Na₂SO₄, filtered and concentrated under vacuum to geta crude residue.

The residue was purified by silica gel column chromatography to afford70 mg of desired product with HPLC purity 35% which was then purified bypreparative HPLC (Conditions: Column: Gemini NX C18 (21.2 mm×150 mmparticle size 5 μm); (Mobile Phase: A; 0.1% Ammonium bicarbonate inWater, B; ACN) to afford the desired compound. The compound was thendiluted with dichloromethane and water. The aqueous layer was separatedand extracted with DCM (3×10 mL). The organic phase was washed withbrine, dried over Na₂SO₄, filtered and concentrated under vacuum toafford the title compound (0.005 g, yield: 11%) as a white solid. 1H-NMR(CD₃OD, 400 MHz): δ 8.41 (s, 1H), 7.79-7.76 (m, 2H), 7.36 (dd, 1H), 6.82(s, 1H), 6.47-6.42 (m, 3H), 5.82 (d, 1H), 3.96 (s, 6H), 3.81-3.55 (m,8H), 2.54 (s, 3H), 2.37 (s, 3H); MS (ESI): 643.1 [M+H]⁺; HPLC: 97.26%,rt: 6.19 min.

Example 149

3-acrylamido-4-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-N,N-dimethylbenzamide

The title compound was synthesized following the approach outlined inProcedure 2K (Example 148), substituting dimethylamine in step (b) toafford the title compound (12.0 mg, yield: 6.1%) as an off-white solid.1H-NMR (CD₃OD, 400 MHz): δ 8.38 (s, 1H), 7.78-7.72 (m, 2H), 7.34 (dd,1H), 6.80 (s, 1H), 6.46-6.38 (m, 3H), 5.80 (dd, 1H), 3.94 (s, 6H), 3.36(s, 3H), 3.10 (s, 6H); MS (ESI): 587.9 [M+H]⁺; HPLC: 99.04%, rt: 3.98min.

Example 150

N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-ethylpiperazine-1-carbonyl)phenyl)acrylamide

The title compound was synthesized following the approach outlined inProcedure 2K (Example 148), substituting 1-ethylpiperazine in step (b)to afford the title compound (10.0 mg, yield: 9.7%) as an off-whitesolid. 1H-NMR (CD₃OD, 400 MHz): δ 8.41 (s, 1H), 7.76-7.79 (m, 2H), 7.36(d, 1H), 6.82 (s, 1H), 6.38-6.47 (m, 3H), 5.82 (d, 1H), 3.96 (s, 6H),3.72-3.82 (m, 2H), 3.53-3.65 (m, 3H), 3.39 (s, 4H), 2.49-2.54 (m, 6H),1.15 (t, 3H); MS (ESI): 657.0 [M+H]⁺; HPLC: 95.98%, rt: 6.25 min.

Example 151

N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-isopropylpiperazin-1-yl)phenyl)acrylamidephosphoric acid

The title compound was synthesized following the approach outlined inProcedure 2J (Example 145), substituting4-(4-isopropylpiperazin-1-yl)-2-nitroaniline (procedure shown below) instep (b) and omitting steps (d) and (e) to afford the free base of thetitle compound (0.1 g, overall yield: 9.7%) as an off-white solid. MS(ESI): 643.1 [M+H]⁺. 85% H₃PO₄ was slowly added to a solution ofN-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-isopropylpiperazin-1-yl)phenyl)acrylamide(0.1 g) in 95% THF-MeOH (5 mL). The resulting reaction mixture was thenallowed to stir at room temperature for 30 min., evaporated the solventand triturated with diethyl ether, dried under vacuum to afford thetitle compound (0.16 g) as a off white solid. ¹HNMR (DMSO-d₆, 400 MHz):δ 12.08 (s, 1H), 9.62 (s, 1H), 8.76 (s, 1H), 8.32 (s, 1H), 7.30 (d, 2H),6.89 (d, 1H), 6.81 (d, 1H), 6.46-6.49 (m, 1H), 6.21-6.25 (m, 2H), 5.72(d, 1H), 3.93 (s, 6H), 3.35-3.41 (m, 1H), 3.23 (s, 3H), 3.12-3.17 (m,4H), 2.67-2.70 (m, 4H), 1.05-1.11 (m, 6H); MS (ESI): 643.3 (M+1); HPLC:95.41%, rt: 6.48 min.

Preparation of 4-(4-isopropylpiperazin-1-yl)-2-nitroaniline

a. tert-butyl 4-isopropylpiperazine-1-carboxylate

Boc piperizine, 1 (10 g, 53.76 mmol) and acetone (4 mL) was taken in amixture of dry DCM (100 mL) and acetic acid (3.2 mL). Stirred at roomtemperature for 20 min. Added Na(OAc)₃BH (17 g, 80.2 mmol) and stirringcontinued at room temperature for 12 hr. The reaction mixture wasdiluted with water, extracted with ethyl acetate (3×25 mL). The organiclayer was washed with brine, dried over Na₂SO₄, filtered andconcentrated under vacuum to get crude title compound (13 g, crude). MS(ESI): 229.2 [M+H]⁺.

b. 1-isopropylpiperazine

TFA (15 mL) was slowly added to a stirred solution of tert-butyl4-isopropylpiperazine-1-carboxylate (13 g crude) in dry DCM (20 mL)under argon atmosphere at 0° C. The resulting reaction mixture wasallowed to warm to room temperature and stirred for 18 hr. The reactionmass was concentrated under vacuum and triturated the residue withn-hexane and diethyl ether. Dried under vacuum to afford the titlecompound (7 g, yield: 97%). MS (ESI): 129.1 [M+H]⁺.

c. tert-butyl (4-(4-isopropylpiperazin-1-yl)-2-nitrophenyl)carbamate

1-isopropylpiperazine (1.29 g, 5.66 mmol) and tert-butyl(4-bromo-2-nitrophenyl)carbamate (1.5 g, 4.71 mmol) were taken in amixture of dry toluene (15 mL) and dioxane (2 mL) in a seal tube underargon atmosphere at room temperature. The Argon gas was purged for 5-10min. Then Cs₂CO₃ (3.06 g, 9.43 mmol) and Xantphos (0.54 g, 0.94 mmol)were added and the resulting reaction mixture was purged with argon gasfor 5 min., followed by Pd₂(dba)₃ (0.43 g, 0.47 mmol). The argon gaspurging was continued for additional 5 min. before sealing the reactionvial. Then the reaction mixture was heated at 100° C. for 12 hr. Aftercompletion of the reaction by TLC, the reaction mixture was cooled toroom temperature, filtered through celite bed and concentrated. Theresidue was purified by silica gel column chromatography (DCM:MeOH/97:3)to afford the title compound (1.2 g, yield: 70.5%). MS (ESI): 365.5[M+H]⁺.

d. 4-(4-isopropylpiperazin-1-yl)-2-nitroaniline

TFA (3 mL) was slowly added to a stirred solution of tert-butyl(4-(4-isopropylpiperazin-1-yl)-2-nitrophenyl)carbamate (1.2 g, 3.29mmol) in dry DCM (5 mL) under argon atmosphere at 0° C. The resultingreaction mixture was allowed to warm to room temperature and stirred for12 hr. Reaction progress was monitored by LCMS, after completion of thereaction, excess solvents were removed under reduced pressure to afforda crude residue. The crude residue was repeatedly washed with ether toget the title compound (1 g, yield: 80.6%) as a red solid. MS (ESI):265.1 [M+H]⁺.

Example—154

Preparation ofN-(5-(4-acetylpiperazin-1-yl)-2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)phenyl)acrylamide

N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(piperazin-1-yl)phenyl)acrylamide(Procedure 2L, Example 157) (8.6 mg, 0.014 mmol) and DIEA (7.5 μl, 0.043mmol) were stirred in DCM (1.0 ml) at room temperature under nitrogenatmosphere. A solution of acetyl chloride (1.0 μl, 0.016 mmol) in DCM(11 ul) was added and the reaction mixture was stirred at roomtemperature for 2 hours. The solvent was evaporated and the resultingmaterial was dissolved in 400 ul of DMSO. The DMSO solution was dilutedwith 1.0 ml of MeOH and purified by prep-HPLC (water/ACN in formic acidcondition) to afford the title compound (1.4 mg, yield: 15%). ¹H-NMR(400 MHz, MeOH-d₄) δ 2.16 (s, 3H) 3.65-3.80 (m, 4H) 3.94 (s, 6H) 5.76(dd, 1H) 6.15 (s, 1H) 6.28-6.49 (m, 2H) 6.78-6.82 (m, 1H) 6.95 (dd, 2.89Hz, 1H) 7.28-7.38 (m, 2H) 8.31 (d, 1H) 8.58 (br. s., 1H); ESI-MS: 643[M+H]⁺.

Example—155 and 156

Preparation ofN-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(1-methoxyethyl)phenyl)acrylamideandN-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(1-hydroxyethyl)phenyl)

The compound was synthesized following the approach outlined inProcedure 2G (Example 123), substituting4-(1-methoxyethyl)-2-nitroaniline (procedure shown below) in step (d) toafford the title compounds after purification using flash chromatographyon silica eluting with 90% to 100% EtOAc/Hexane. Non-polar fraction wasN-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(1-methoxyethyl)phenyl)acrylamide(16 mg, yield: 6% in five steps)¹H-NMR (400 MHz, MeOH-d₄) δ 1.46 (d, 3H)3.29 (s, 3H) 3.96 (s, 6H) 4.12 (d, 1H) 4.40 (q, 1H) 5.78-5.82 (m, 1H)6.33-6.50 (m, 3H) 6.82 (s, 1H) 7.27 (dd, 1H) 7.55-7.64 (m, 2H) 8.37 (d,1H); ESI-MS: 575 [M+H]⁺. Polar fraction wasN-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(1-hydroxyethyl)phenyl)acrylamide (22 mg, yield: 8% in five steps). ¹H-NMR (400 MHz, MeOH-d₄) δ1.53 (d, 3H) 4.00 (s, 6H) 5.83 (dd, 1H) 6.29-6.56 (m, 3H) 6.86 (s, 1H)7.38 (dd, 1H) 7.57 (d, 1H) 7.69 (s, 1H) 8.40 (d, 1H); ESI-MS: 661[M+H]⁺.

Preparation of 4-(1-methoxyethyl)-2-nitroaniline

a. 1-(4-fluoro-3-nitrophenyl)ethanol

1-(4-fluoro-3-nitrophenyl)ethanone (1.0 g, 5.46 mmol) was dissolved inMeOH (15.0 ml) and stirred over ice bath. NaBH₄ (0.62 g, 16.0 mmol) wasadded portionwise. Upon completion of the addition, the reaction mixturewas stirred at rt for 10 minutes. The reaction mixture was poured intoEtOAc and brine. The organic layer was separated, dried over MgSO₄ andevaporated. The resulting material was purified by flash chromatographyon silica eluting with 20% to 70% EtOAc/Hexane to afford the titlecompound (881 mg, yield: 87%). %). ¹H-NMR (400 MHz, CDCl₃) δ 1.53 (d,4H) 4.99 (dd, 1H) 7.22-7.33 (m, 1H) 7.66 (ddd, 1H) 8.09 (dd, 1H)

b. 1-fluoro-4-(1-methoxyethyl)-2-nitrobenzene

1-(4-fluoro-3-nitrophenyl)ethanol (870 mg, 4.7 mmol) was dissolved inMeOH (10 ml) and concentrated sulfuric acid (2.5 ml, 47 mmol) was addedcarefully. The reaction mixture was heated at 150° C. using microwave(Biotage Initiator) for 5 minutes. After cooling to room temperature,the reaction mixture was poured into EtOAc/water. The organic layer waswashed with brine, dried over MgSO₄ and evaporated. The resultingmaterial was purified by flash chromatography on silica eluting with 0%to 50% EtOAc/Hexane to afford the title compound (664 mg, yield: 71%).¹H-NMR (400 MHz, CDCl₃) δ 1.45 (d, 3H) 3.27 (s, 3H) 4.36 (q, 1H)7.28-7.32 (m, 1H) 7.56-7.63 (m, 1H) 7.98-8.04 (m, 1H)

c. 4-(1-methoxyethyl)-2-nitroaniline

1-fluoro-4-(1-methoxyethyl)-2-nitrobenzene (664 mg, 3.33 mmol) wasstirred in THF (10 ml). NH₄OH (0.39 ml, 10.0 mmol) was added and thereaction mixture was stirred at room temperature for 2 hours. Further780 ul of NH₄OH was added and the reaction mixture was heated at 120° C.for 10 minutes using microwave (Biotage Initiator) and then at 140° C.for 50 minutes. After cooling to room temperature, the reaction mixturewas poured into EtOAc/water. The organic layer was washed with brine,dried over MgSO₄ and evaporated. The resulting material was purified byflash chromatography on silica eluting with 0% to 50% EtOAc/Hexane toafford the title compound (290 mg, yield: 44%). ¹H-NMR (400 MHz, CDCl₃)δ 1.42 (d, 3H) 3.22 (s, 3H) 4.24 (q, 1H) 6.05 (br. s., 2H) 6.83 (d, 1H)7.38 (dd, 2.01 Hz, 1H) 8.04 (d, 1H)

Procedure 2L: Example—157

Preparation ofN-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(piperazin-1-yl)phenyl)acrylamide

a. 2-(4-bromo-2-nitrophenyl)isoindoline-1,3-dione

4-bromo-1-fluoro-2-nitrobenzene (2.0 g, 9.1 mmol) and potassium1,3-dioxoisoindolin-2-ide (2.0 g, 10.9 mmol) were placed in microwavevial (0.5-2 ml) and NMP (12.0 ml) was added. The reaction mixture washeated at 200° C. using microwave (Biotage, Initiator) for 1 min. Aftercooling to rt, the reaction mixture was added dropwise to stirring waterwhich resulted in formation of precipitate. The solid was collected,washed with water and dried under stream of nitrogen to afford the titlecompound (2.89 g, yield: 92%). ¹H-NMR (400 MHz, CDCl₃) δ 7.26 (s, 2H)7.43 (d, 2H) 7.81-7.87 (m, 3H) 7.91 (dd, 2H) 7.95-8.05 (m, 3H) 8.33 (d,1H)

b. tert-butyl4-(4-(1,3-dioxoisoindolin-2-yl)-3-nitrophenyl)piperazine-1-carboxylate

2-(4-bromo-2-nitrophenyl)isoindoline-1,3-dione (500 mg, 1.4 mmol),tert-butyl piperazine-1-carboxylate (402 mg, 2.2 mmol), Pd₂(dba)₃ (66.0mg, 0.07 mmol), Xantphos (83 mg, 0.144 mmol) and Cs₂CO₃ (939 mg, 2.9mmol) were placed in reaction vial (microwave reaction vial 10-20 ml)and purged with nitrogen. Toluene (5.0 ml) was added and nitrogen wasbubbled for 10 min. The reaction mixture was heated at 100° C. for 5hours. Heating was stopped and cooled to rt. The reaction mixture wasfiltered through a pad of celite and filtrate was evaporated. Theremaining material was purified by flash chromatography on silicaeluting with 20% to 70% EtOAc/Hexane to afford the title compound (275mg, yield: 42%). ¹H-NMR (400 MHz, CDCl₃) δ 1.51 (s, 6H) 3.28-3.39 (m,2H) 3.57-3.70 (m, 2H) 7.20-7.24 (m, 1H) 7.35 (d, 1H) 7.66 (d, 1H)7.75-7.87 (m, 1H) 7.96 (dd, 1H)

c. tert-butyl 4-(4-amino-3-nitrophenyl)piperazine-1-carboxylate

To a suspension of tert-butyl4-(4-(1,3-dioxoisoindolin-2-yl)-3-nitrophenyl)piperazine-1-carboxylate(275.4 mg, 0.6 mmol) in THF (5.0 ml) was added 1.0M Hydrazine in THF(1.8 ml, 1.8 mmol) at room temperature. The reaction mixture was stirredat room temperature for 2 hours. The reaction mixture was poured intoEtOAc/water and organic layer was washed with brine, dried over MgSO₄and evaporated. The resulting oil was purified by flash chromatographyon silica eluting with 20% to 70% EtOAc/Hexane to afford the titlecompound (182 mg, 93% yield). ¹H-NMR (400 MHz, DMSO-d₆) δ 1.42 (s, 9H)2.89-2.99 (m, 4H) 3.44 (d, 4H) 6.98 (d, 1H) 7.22 (s, 2H) 7.26-7.38 (m,2H)

d. tert-buty4-(4-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-((2-(trimethylsilyl)ethoxy)methy)ureido)pyrimidin-4-yl)amino)-3-nitrophenyl)piperazine-1-carboxylate

1-(6-chloropyrimidin-4-yl)-3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-((2-(trimethylsilyl)ethoxy)methyl)urea(175 mg, 0.34 mmol), tert-butyl4-(4-amino-3-nitrophenyl)piperazine-1-carboxylate (90 mg, 0.28 mmol),Pd₂(dba)₃ (13 mg, 0.014 mmol), Brettphos (13 mg, 0.028 mmol) and sodiumtert-butoxide (54 mg, 0.56 mmol) were placed in a reaction vial (2 to 5ml) and purged with nitrogen. Toluene (1.0 ml) was added and nitrogenwas bubbled for 5 min then the reaction mixture was heated at 100° C.overnight. Heating was stopped and cooled to room temperature. Thereaction mixture was filtered through pad of Celite®. The filtrate wasevaporated and resulting material was purified by flash chromatographyon silica eluting with 20% to 70% EtOAc/Hexane to afford the titlecompound (102.4 mg, yield: 45%). ¹H-NMR (400 MHz, CDCl₃) δ 0.01 (s, 9H)0.85-0.98 (m, 2H) 1.50 (s, 9H) 3.03 (s, 3H) 3.17 (br. s., 4H) 3.56-3.66(m, 4H) 3.75-3.97 (m, 8H) 5.22 (s, 2H) 6.50 (s, 1H) 6.95 (s, 1H) 7.65(s, 1H) 8.46 (s, 1H)

e. tert-butyl4-(4-((tert-butoxycarbonyl)(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-((2-(trimethylsilyl)ethoxy)methyl)ureido)pyrimidin-4-yl)amino)-3-nitrophenyl)piperazine-1-carboxylate

tert-butyl4-(4-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-((2-(trimethylsilyl)ethoxy)methyl)ureido)pyrimidin-4-yl)amino)-3-nitrophenyl)piperazine-1-carboxylate(162 mg, 0.2 mmol), di-tert-butyl dicarbonate (53 mg, 0.24 mmol) andDMAP (4.9 mg, 0.04 mmol) were stirred in THF (2.0 ml) at roomtemperature under nitrogen atmosphere for 1 hour. The reaction mixturewas poured into EtOAc and water. The organic layer was washed withbrine, dried over MgSO₄ and evaporated. The resulting material waspurified by flash chromatography on silica eluting with 10% to 70%EtOAc/Hexane to afford the title compound (148 mg, yield: 82%). ¹H-NMR(400 MHz, CDCl₃) δ 0.01 (s, 9H) 0.80-1.04 (m, 2H) 1.4 (s, 9H) 1.49 (s,9H) 3.12 (s, 3H) 3.19-3.36 (m, 4H) 3.50-3.70 (m, 4H) 3.83-3.92 (m, 9H)3.96 (d, 11H) 5.14 (d, 1H) 5.42 (d, 1H) 6.46 (s, 1H) 7.05-7.19 (m, 2H)7.58 (d, 1H) 8.03 (s, 1H) 8.29 (s, 1H)

f. tert-butyl 4-(3-amino-4-((tert-butoxycarbonyl)(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-J-methyl-3-((2-(trimethylsilyl)ethoxy)methyl)ureido)pyrimidin-4-yl)amino)phenyl)piperazine-1-caroxylate

tert-butyl4-(4-((tert-butoxycarbonyl)(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-((2-(trimethylsilyl)ethoxy)methyl)ureido)pyrimidin-4-yl)amino)-3-nitrophenyl)piperazine-1-carboxylate(149 mg, 0.164 mmol) was stirred in THF (1.5 ml) and MeOH (1.5 ml). Fivedrops of Raney Nickel suspension in water was added. The solution wasstirred under hydrogen atmosphere at room temperature overnight. Thereaction was filtered through a pad of Celite®. and filtrate wasconcentrated to give crude title compound which was taken to the nextstep without further purification. ESI-MS: 877 [M+H]⁺.

g. tert-butyl 4-(3-acrylamido-4-((tert-butoxycarbonyl)(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-((2-(trimethylsilyl)ethoxy)methyl)ureido)pyrimidin-4-yl)amino)phenyl)piperazine-1-carboxylate

Crude tert-butyl4-(3-amino-4-((tert-butoxycarbonyl)(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-((2-(trimethylsilyl)ethoxy)methyl)ureido)pyrimidin-4-yl)amino)phenyl)piperazine-1-carboxylate(95 mg, 0.11 mmol) was dissolved in THF (1.5 ml) and stirred over icebath under nitrogen atmosphere. DIEA (57 μl, 0.30 mmol) followed byacryloyl chloride (13 μl, 0.16 mmol) were added and the reaction mixturewas stirred at room temperature for 1 hour. The reaction mixture waspoured into EtOAc and brine. The organic. layer was separated, driedover MgSO₄ and evaporated. The resulting material was purified by flashchromatography on silica eluting with 30% to 100% EtOAc/Hexane to affordthe title compound (47 mg, yield: 47%). ¹H-NMR (400 MHz, CDCl₃) δ 0.02(s, 9H) 0.79-1.04 (m, 2H) 1.36 (s, 9H) 1.49 (s, 9H) 3.08 (s, 4H) 3.05(s, 3H) 3.25 (br. s., 4H) 3.64 (br. s., 4H) 3.86 (s, 6H) 5.27 (s, 1H)5.67-5.79 (m, 1H) 6.15-6.56 (m, 2 II) 6.48 (s, 1H) 7.03-7.14 (m, 1H)7.87-8.09 (m, 2H) 8.19-8.29 (br. s., 1H) 8.44 (s, 1H).

h.N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(piperazin-1-yl)phenyl)acrylamide

tert-butyl4-(3-acrylamido-4-((tert-butoxycarbonyl)(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-((2-(trimethylsilyl)ethoxy)methyl)ureido)pyrimidin-4-yl)amino)phenyl)piperazine-1-carboxylate(59 mg, 0.06 mmol) was stirred in DCM (2.0 ml). TFA (97 μl, 1.3 mmol)was added and the mixture was stirred at room temperature for 6 hours.The reaction mixture was evaporated. The remaining residue was dissolvedin DCM and washed with saturated NaHCO₃, dried over MgSO₄ andevaporated. The resulting material was dissolved in THF (2.0 ml) andNH₄OH (74 μl, 1.9 mmol) was added. The reaction mixture was stirred atroom temperature for 5 minutes. The reaction mixture was evaporated toobtain the title compound (34 mg, yield: 90%) which was taken to thenext step without further purification. ¹H-NMR (400 MHz, DMSO-d₆) δ 2.62(br. s., 4H) 3.08-3.27 (m, 7H) 3.77-4.03 (m, 6H) 5.72 (d, 1H) 6.11-6.34(m, 2H) 6.40-6.62 (m, 1H) 6.70-6.98 (m, 2H) 7.22-7.37 (m, 2H) 8.32 (s,1H) 8.70 (s, 1H) 9.58 (br. s., 1H) 12.06 (s, 1H); ESI-MS: 601 [M+H]⁺.

Example—158

N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-((2-hydroxyethyl)(methyl)amino)phenyl)acrylamide

The compound was synthesized following the approach outlined inProcedure 2L (Example 157), substituting2-((tert-butyldimethylsilyl)oxy)-N-methylethanamine, Pd(dba)₂ and Ruphosin step (b), and Pd(dba)₂, Brettphos and sodium tert-butoxide in step(d) to afford the title compound (64 mg, yield: 16% in seven steps)¹H-NMR (400 MHz, MeOH-d₄) δ 3.04 (s, 3H) 3.26 (s, 3H) 3.51 (s, 2H)3.69-3.81 (m, 2H) 3.94 (s, 6H) 5.71-5.78 (m, 1H) 6.03-6.10 (m, 1H)6.26-6.45 (m, 2H) 6.72 (d, 1H) 6.79 (s, 1H) 7.04-7.10 (m, 1H) 7.23 (d,1H) 8.28 (d, 1H); ESI-MS: 590 [M+H]⁺.

Preparation of 2-((tert-butyldimethylsilyl)oxy)-N-methylethanamine

a. 2-((tert-butyldimethylsilyl)oxy)-N-methylethanamine

2-(methylamino)ethanol (1.0 g, 13.3 mmol) was stirred in DCM (25.0 ml)under atmosphere of nitrogen. DIEA (3.23 ml, 18.6 mmol) followed bytert-butylchlorodimethylsilane (2.0 g, 13.3 mmol) was added and thereaction mixture was stirred at room temperature over night. Thereaction mixture was poured into ether/water. The aqueous layer wasextracted with ether three times. The combined organic layer was driedover MgSO₄ and evaporated and remaining material was dried under highvacuum to afford title compound (1.4 g, yield: 56%). ¹H-NMR (400 MHz,CDCl₃) δ 0.08 (s, 6H) 0.91 (s, 9H) 1.93 (br. s., 1H) 2.48 (s, 3H) 2.71(t, 2H) 3.75 (t, 2H).

Example—160

N-(5-(1-aminoethyl)-2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)phenyl)acrylamideTFA salt

The compound was synthesized following the approach outlined inProcedure 2G (Example 123) substituting tert-butyl(1-(4-amino-3-nitrophenyl)ethyl)carbamate, Pd(dba)₂, Brettphos andsodium tert-butoxide in step (d) to afford the free base which wasconverted to TFA salt. The free base was dissolved in DCM and 1equivalent of TFA was added. The mixture was evaporated and triturationwith ether afforded the title compound (65 mg, yield: 23% in fivesteps). ¹H-NMR (400 MHz, MeOH-d₄) δ 1.67 (d, 3 II) 3.33-3.37 (s, 3H)3.92-3.97 (s, 6H) 4.49 (m, 1H) 5.75-5.83 (m, 1 II) 6.30-6.50 (m, 3H)6.81 (s, 1H) 7.36 (dd, 1H) 7.69 (d, 1H) 7.82 (d, 1H) 8.36-8.43 (m, 1H);ESI-MS: 560 [M+H]⁺.

Preparation of tert-butyl (1-(4-amino-3-nitrophenyl)ethyl)carbamate

a. 1-(4-fluoro-3-nitrophenyl)ethanamine

Fuming HNO₃ (0.48 ml, 10.8 mmol) was added carefully to ice bath cooledconcentrated H₂SO₄ (3.6 ml, 68.2 mmol). To the mixture was added1-(4-fluorophenyl)ethanamine (1.0 g, 7.2 mmol) dropwise. The reactionmixture was stirred with cooling for 50 minutes. The reaction mixturewas poured into ice and basified with 3M NaOH solution (24 ml, 72.00mmol) to about pH8.0. The alkaline solution was extracted with DCMtwice. The combined organic layer was dried over Na₂SO₄ and evaporatedto afford title compound (1.3 g, yield: 96%). ¹H-NMR (400 MHz, CDCl₃) δ1.41 (d, 6H) 4.25 (q, 1H) 7.21-7.26 (m, 1H) 7.67 (ddd, 1H) 8.10 (dd, 1H)

b. tert-butyl (1-(4-fluoro-3-nitrophenyl)ethyl)carbamate

To the suspension of 1-(4-fluoro-3-nitrophenyl)ethanamine (1.3 g, 6.9mmol) in THF (10.0 ml) was added DIEA (2.4 ml, 13.8 mmol) followed bydi-tert-butyl dicarbonate (1.9 ml, 8.3 mmol). The reaction mixture wasstirred at room temperature under nitrogen atmosphere for 2 hours. Thereaction mixture was poured into EtOAc/water and organic layer waswashed with brine, dried over MgSO₄ and evaporated. The resultingmaterial was purified by flash chromatography on silica eluting with 10%to 70% EtOAc/Hexane to afford the title compound (1.5 g, yield: 75%).¹H-NMR (400 MHz, CDCl₃) δ 1.37-1.50 (m, 12H) 4.81 (br. s., 2H) 7.23-7.26(m, 1H) 7.59 (ddd, 1H) 8.01 (dd, 1H).

c. tert-butyl (1-(4-amino-3-nitrophenyl)ethyl)carbamate

Tert-butyl (1-(4-fluoro-3-nitrophenyl)ethyl)carbamate (500 mg, 1.75mmol) was stirred in THF (2 ml). NH₄OH (0.978 ml, 7.0 mmol) was addedand the reaction mixture was heated at 150° C. for 30 minutes usingmicrowave (Biotage Initiator). NH₄OH (0.600 ml) was added and thenheated at 180° C. After cooling to room temperature, the reactionmixture was poured into EtOAc/water. The organic layer was washed withbrine, dried over MgSO₄ and evaporated. The resulting material waspurified by flash chromatography on silica eluting with 0% to 50%EtOAc/Hexane to afford the title compound (376 mg, yield: 76%) ¹H-NMR(400 MHz, CDCl₃) δ 1.43-1.48 (m, 12H) 4.71 (br. s., 2H) 6.79 (d, 1H)7.34 (dd, 1H) 8.05 (d, 1H).

Example—161

Preparation ofN-(5-(4-(2-aminoethyl)piperazin-1-yl)-2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)phenyl)acrylamide2 TFA salt

a. tert-butyl(2-(4-(3-acrylamido-4-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)phenyl)piperazin-1-yl)ethyl)carbamate

To the solution ofN-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(piperazin-1-yl)phenyl)acrylamide2,2,2-trifluoroacetate (Procedure 2L, Example 157) (30 mg, 0.042 mmol)in THF (1.0 ml) and MeOH (1.0 ml) was added tert-butyl(2-oxoethyl)carbamate (13 mg, 0.08 mmol). The reaction mixture wasstirred at room temperature for 10 minutes. Sodium cyanoborohydride (7.0mg, 0.12 mmol) was added and the reaction mixture was stirred at roomtemperature over night. Solvent was evaporated and saturated NaHCO₃solution was added. The mixture was extracted three times with DCM.Combined organic layer was dried over Na₂SO₄ and evaporated. Theremaining material was purified by flash chromatography on silicaeluting with 0% to 15% MeOH in DCM to afford the title compound (17 mg,yield: 54%) ESI-MS: 744 [M+H]⁺.

N-(5-(4-(2-aminoethyl)piperazin-1-yl)-2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)phenyl)acrylamide2 TFA salt

tert-butyl(2-acrylamido-4-(4-(2-((tert-butoxycarbonyl)amino)ethyl)piperazin-1-yl)phenyl)(6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-((2-(trimethylsilyl)ethoxy)methyl)ureido)pyrimidin-4-yl)carbamate (17 mg, 0.017 mmol) was stirred in DCM (1.0ml). TFA (200 μl, 2.6 mmol) was added and the mixture was stirred atroom temperature for 2 hours. Solvent was evaporated and remainingmaterial was triturated with ether. The resulting solid was collectedwashed with ether and dried under stream of nitrogen to afford the titlecompound (15 mg, yield: 96%). ¹H-NMR (400 MHz, MeOH-d₄) δ 2.90 (br. s.,6H) 3.15-3.22 (m, 2H) 3.33-3.41 (m, 4H) 3.94 (s, 6H) 5.74-5.79 (m, 1H)6.17 (s, 1H) 6.32-6.44 (m, 2H) 6.81 (s, 1H) 6.92-6.98 (m, 1H) 7.35 (d,2H) 8.32 (d, 1H); ESI-MS: 644 [M+H]⁺.

Example—162

N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-3-fluorophenyl)acrylamide2,2,2-trifluoroacetate

The compound was synthesized following the approach outlined inProcedure 2G (Example 123), substituting 2-fluoro-6-nitroaniline,Pd(dba)₂, Brettphos and sodium tert-butoxide in step (d) to afford thefree base, which was converted to TFA salt. The free base was dissolvedin DCM and 1 equivalent of TFA was added. The mixture was evaporated andtrituration with ether afforded the title compound (47 mg, yield: 22% infour steps). ¹H-NMR (400 MHz, MeOH-d₄) δ 3.30 (s, 3H) 3.93 (s, 6H)5.72-5.78 (m, 2H) 6.21-6.29 (m, 1H) 6.23 (d, 1H) 6.27 (d, 1H) 6.32-6.40(m, 1H) 6.56 (dd, 1H) 6.90 (s, 1H) 7.10 (t, 1H) 7.25-7.37 (m, 1H) 7.76(d, 1H) 8.32 (s, 1H) 8.89 (s, 1H) 9.66 (s, 1H) 11.96 (s, 1H); ESI-MS:535 [M+H]⁺

Example—163

N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(2-(dimethylamino)ethyl)phenyl)acrylamide2,2,2-trifluoroacetate

The compound was synthesized following the approach outlined inProcedure 2G (Example 123), substituting tert-butyl4-amino-3-nitrophenethylcarbamate (procedure shown below), Pd(dba)₂ andBrettphos in step (d) to afford crude TFA salt of the free terminalamine, which was used as-is in the following additional step: To thesolution ofN-(5-(2-aminoethyl)-2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-3-(hydroxymethyl)-1-methylureido)pyrimidin-4-yl)amino)phenyl)acrylamide2,2,2-trifluoroacetate (9.2 mg, 0.014 mmol) in THF (1.0 ml) and MeOH(1.0 ml) was added formaldehyde (5.1 μl, 0.068 mmol). The reactionmixture was stirred at room temperature for 10 minutes. Sodiumcyanoborohydride (3.4 mg, 0.055 mmol) was added and the reaction mixturewas stirred at room temperature over night. Solvent was evaporated andsaturated NaHCO₃ solution was added. The mixture was extracted threetimes with DCM. Combined organic layer was dried over Na₂SO₄ andevaporated. The remaining material was purified by flash chromatographyon silica eluting with 10% to 70% MeOH in DCM to afford free base. To asolution of free base in DCM, TFA (11 μl, 0.014 mmol) was added. Thesolvent was evaporated and dried under high vacuum to afford the titlecompound (6.3 mg, yield: 66%). ¹H-NMR (400 MHz, MeOH-d₄) δ 2.97 (s, 6H)3.10 (dd, 2H) 3.34 (s, 3H) 3.45 (dd, 2H) 3.94 (s, 6H) 4.63-4.63 (m, 1H)5.76-5.81 (m, 1H) 6.31-6.48 (m, 3H) 6.81 (s, 1H) 7.25 (dd, 1H) 7.55 (d,1H) 7.62-7.72 (m, 1H) 8.35 (s, 1H); ESI-MS: 588 [M+H]⁺.

Preparation of tert-butyl 4-amino-3-nitrophenethylcarbamate

a. tert-butyl 4-fluoro-3-nitrophenethylcarbamate

2-(4-fluorophenyl)ethanamine (1.0 g, 7.2 mmol) was dissolved inconcentrated H₂SO₄ (4.0 ml, 75 mmol) and cooled over ice bath. FumingHNO₃ (0.48 ml, 10.8 mmol) was added carefully dropwise to the mixture.The reaction mixture was stirred with cooling for 45 minutes and pouredinto ice. The mixture was basified with 3M NaOH solution (60 ml, 180mmol) and the alkaline solution was extracted with DCM three times. Thecombined organic layer was dried over Na₂SO₄ and evaporated to affordcrude amine. The crude material was dissolved in THF (15.0 ml).Di-tert-butyl dicarbonate (1.7 g, 7.9 mmol) and DIEA (2.5 ml, 14.4 mmol)were added and the reaction mixture was stirred at room temperature for12 hours. The reaction mixture was poured into water and extracted withEtOAc. The organic layer was washed with brine, dried over MgSO₄ andevaporated. The resulting material was purified by flash chromatographyon silica eluting with 0% to 20% EtOAc/Hexane to afford title compound(569 mg, yield: 28%). ¹H-NMR (400 MHz, CDCl₃) δ 1.44 (s, 9H) 2.88 (t,2H) 3.40 (d, 2H) 7.24 (t, 1H) 7.42-7.54 (m, 1H) 7.89 (dd, 1H)

b. tert-butyl 4-amino-3-nitrophenethylcarbamate

Tert-butyl 4-fluoro-3-nitrophenethylcarbamate (569 mg, 2.00 mmol) wasstirred in THF (2.64 ml). NH₄OH (2.50 ml, 17.97 mmol) was added and thereaction mixture was heated at 150° C. for 30 minutes using microwave(Biotage Initiator). NH₄OH (0.600 ml) was added and then heated at 180°C. After cooling to room temperature, the reaction mixture was pouredinto EtOAc/water. The organic layer was washed with brine, dried overMgSO₄ and evaporated. The resulting material was purified by flashchromatography on silica eluting with 0% to 30% EtOAc/Hexane to affordthe title compound (257 mg, yield: 46%) ¹H-NMR (400 MHz, CDCl₃) δ 1.44(s, 9H) 2.73 (t, J=7.03 Hz, 2H) 3.34 (m, 2H) 4.46-4.62 (m, 1H) 6.78 (d,J=8.53 Hz, 1H) 7.24 (dd, J=8.53, 1.76 Hz, 1H) 7.94 (d, J=1.76 Hz, 1H).

Example—164

N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(2-(dimethylamino)ethyl)phenyl)acrylamide2,2,2-trifluoroacetate

The compound was synthesized following the approach outlined inProcedure 2L (Example 157), substituting2-nitro-4-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)aniline (preparationshown below) and sodium tert-butoxide in step (d) to afford the titlecompound (7 mg, yield: 2.7% over four steps), ¹H-NMR (400 MHz, MeOH-d₄)δ 3.28 (s, 3H) 3.88-3.97 (m, 8H) 4.06-4.14 (m, 2H) 5.76 (dd, 1H) 6.12(s, 1H) 6.32-6.46 (m, 2H) 6.79 (s, 1H) 6.91 (dd, 1H) 7.34-7.41 (m, 2H)8.31 (d, 1H); ESI-MS: 577 [M+H]⁺.

Preparation of2-nitro-4-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)aniline

a. 2-nitro-4-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)aniline

4-amino-3-nitrophenol (2.0 g, 12.977 mmol) was dissolved in DMF (20 ml)and Potassium carbonate (3.59 g, 25.953 mmol) was added. To the mixturewas added 2-(2-bromoethoxy)tetrahydro-2H-pyran (2.55 ml, 16.87 mmol) andthe reaction mixture was stirred at 50° C. for 6 hours and then for 3days at room temperature. The reaction mixture was poured intoEtOAc/brine. The aqueous layer was extracted twice with EtOAc. Thecombined organic layer was washed with brine three times, dried overMgSO4 and evaporated. The resulting material was purified by flashchromatography on silica eluting with 5% to 40% EtOAc/Hexane to affordtitle compound (1.4 g, yield: 38%). ¹H-NMR (400 MHz, CDCl₃) δ 1.51-1.68(m, 4H) 1.71-1.89 (m, 2H) 3.46-3.61 (m, 1H) 3.72-3.99 (m, 2H) 4.03-4.24(m, 3H) 4.64-4.79 (m, 1H) 6.78 (d, 1H) 7.09-7.17 (m, 1H) 7.60 (d, 1H).

Example—165

N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-3-methylphenyl)acrylamide

The compound was synthesized following the approach outlined inProcedure 2G (Example 123), substituting 2-methyl-6-nitroaniline,Pd(dba)₂, Brettphos and sodium tert-butoxide in step (d) to afford thetitle compound (3.0 mg, yield: 6% in five steps). ¹H-NMR (400 MHz,MeOH-d₄) δ 3.52 (m, 3H) 3.95 (m, 6H) 5.69 (dd, 1H) 6.21 (dd, 1H) 6.46(dd, 1H) 6.64 (d, 1H) 6.73 (d, 1H) 6.82 (s, 1H) 7.11 (t, 1H) 8.06 (d,1H) 8.55 (d, 1H); ESI-MS: 531 [M+H]

Example—166

N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-3-methoxyphenyl)acrylamide

The compound was synthesized following the approach outlined inProcedure 2G (Example 123), substituting 2-methoxy-6-nitroaniline,Pd(dba)₂, Brettphos and sodium tert-butoxide in step (d) to afford thetitle compound (33.0 mg, yield: 6.1% in five steps). ¹H-NMR (400 MHz,MeOH-d₄) δ 3.76 (s, 3H) 3.93 (s, 6H) 5.70 (dd, 1H) 6.23 (d, 1H) 6.52 (d,1H) 6.88-6.95 (m, 2H) 7.27 (t, 1H) 7.44-7.56 (m, 1H) 8.28 (s, 1H) 8.57(br. s., 1H) 9.40-9.57 (m, 1H) 12.15 (s, 1H); ESI-MS: 547 [M+H]⁺

Example—167

N-(3-chloro-2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)phenyl)acrylamide

The compound was synthesized following the approach outlined inProcedure 2G (Example 123), substituting 2-chloro-6-nitroaniline,Pd(dba)₂, Brettphos and sodium tert-butoxide in step (d) to afford thetitle compound (11.0 mg, yield: 5% in five steps). ¹H-NMR (400 MHz,MeOH-d₄) δ 3.29 (s, 3H) 3.94 (s, 6H) 5.71-5.75 (m, 1H) 6.23 (dd, 1H)6.56 (dd, 1H) 6.90 (s, 1H) 7.31-7.39 (m, 2H) 7.94 (d, 1H) 8.30 (s, 1H)9.01 (s, 1H) 9.51-9.70 (m, 1H) 12.04 (s, 1H); ESI-MS: 551 [M+H]⁺.

Example—168

Preparation ofN-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-(2-hydroxyethyl)piperazin-1-yl)phenyl)acrylamideTFA salt

The compound was synthesized following the approach outlined in Example161, substituting 2-((tert-butyldimethylsilyl)oxy)acetaldehyde in step(a) to afford the title compound (13 mg, yield: 42% two steps)¹H-NMR(400 MHz, MeOH-d₄) δ 3.14-3.20 (m 2H) 3.35-3.37 (m, 2H) 3.73-3.76 (m,2H) 3.84-3.96 (m, 10H) 5.75-5.79 (m, 1H) 6.24 (s, 1H) 6.37-6.47 (m, 2H)6.81 (s, 1H) 6.99 (dd, 1H) 7.37-7.46 (m, 2H) 8.34-8.36 (m, 1H); ESI-MS:645 [M+H]⁺.

Example—170

(E)-3-chloro-N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)phenyl)acrylamide

The title compound was synthesized following the approach outlined inProcedure 2A (Example 100), modifying step (i) to the followingprocedure: To a solution of 1-(6-((2-aminophenyl)amino)pyrimidin-4-yl)-3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylurea (10mg, 0.022 mmol), triethylamine (10.9 mg, 0.11 mmol), and(E)-3-chloroacrylic acid (2.76 mg, 0.026 mmol) in DCM (0.4 ml, 6.22mmol), cooled to 0° C., was added2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphinane 2,4,6-trioxide (0.023ml, 0.039 mmol, 50% solution in EtOAc). The resulting mixture wasstirred at room temperature for 4 h and concentrated. The remainingresidue was purified by silica gel flash chromatography to obtain thetitle compound (7.2 mg, yield: 61%). ¹HNMR (400 MHz, CDCl₃) δ 3.30 (s,3H) 3.90 (s, 6H) 6.02 (s, 1H) 6.36 (d, J=12.92 Hz, 1H) 6.49 (s, 1H)7.21-7.33 (m, 2H) 7.36-7.50 (m, 2H) 7.75 (d, J=6.53 Hz, 1H) 7.93 (br.s., 1H) 8.41 (s, 1H) 12.50 (s, 1H); MS (ESI): 551.0 [M+H]⁺

Example—171

(Z)-3-chloro-N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)phenyl)acrylamide

The title compound was synthesized following the approach outlined inProcedure 2A (Example 100), modifying step (i) to the followingprocedure: To a solution of 1-(6-((2-aminophenyl)amino)pyrimidin-4-yl)-3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylurea (20mg, 0.043 mmol), triethylamine (21.8 mg, 0.22 mmol), and(Z)-3-chloroacrylic acid (5.5 mg, 0.052 mmol) in DCM (0.86 ml, 13.3mmol), cooled to 0° C. was added2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphinane 2,4,6-trioxide (49 mg,0.78 mmmol, 50% solution of EtOAc). The resulting mixture was stirred atroom temperature for 2 h and concentrated. The residue was purified bysilica gel column chromatography to obtain the title compound (15 mg,yield: 63%). ¹H NMR (400 MHz, CDCl₃) δ X; MS (ESI): 551.0 [M+H]⁺.

Example—172

(Z)—N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)phenyl)but-2-enamide

The title compound was synthesized following the approach outlined inProcedure 2A (Example 100), modifying step (i) to the followingprocedure: To a solution of 1-(6-((2-aminophenyl)amino)pyrimidin-4-yl)-3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylurea (20mg, 0.043 mmol), triethylamine (21.8 mg, 0.22 mmol), and (Z)-but-2-enoicacid (4.5 mg, 0.052 mmol) in DCM (0.86 ml, 13.3 mmol) was added asolution of 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphinane2,4,6-trioxide (49.4 mg, 0.78 mmol, 50% solution EtOAc) at 0° C., andthe resulting mixture was stirred at room temperature for 2 h. Themixture was concentrated and the residue was purified by silica gelcolumn to obtain the title compound (8.5 mg, yield: 37%). 1H NMR (400MHz, CDCl₃) δ 2.20 (dd, J=7.28, 1.76 Hz, 3H) 3.30 (s, 3H) 3.92 (s, 6H)5.85 (dd, J=11.42, 1.76 Hz, 1H) 5.99 (s, 1H) 6.29 (dd, J=11.36, 7.34 Hz,1H) 6.52 (s, 1H) 7.23-7.34 (m, 2H) 7.46 (d, J=7.40 Hz, 1H) 7.55 (br. s.,1H) 7.78 (d, J=7.53 Hz, 1H) 8.40 (d, J=0.88 Hz, 1H) 12.50 (s, 1H); MS(ESI): 531.3 [M+H]⁺.

Example—175

(S,Z)—N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)phenyl)-4-hydroxypent-2-enamide

The compound was synthesized following the approach outlined inProcedure 2G (Example 123), substituting 2-nitroaniline in step (d) andmethacryloyl chloride in step (g) to afford the title compound (13 mg,yield: 72%). ¹H NMR (400 MHz, CDCl₃) 2.03 (br. s., 3H) 3.31 (s, 3H) 3.92(s, 6H) 5.49 (s, 1H) 5.81 (s, 1H) 6.00 (s, 1H) 6.53 (s, 1H) 7.29 (m, 3H)7.44 (d, J=7.91 Hz, 1H) 7.83 (d, J=7.53 Hz, 1H) 7.92 (s, 1H) 8.41 (s,1H) 12.34 (s, 1H). MS (ESI): 531.1 [M+H]⁺.

Example—181

(Z)-3-chloro-N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-5-(4-ethylpiperazin-1-yl)phenyl)acrylamide

The title compound was synthesized following the approach outlined inProcedure 2C (Example 108), modifying step (g) to the followingprocedure: To a solution of1-(6-((2-amino-4-(4-ethylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)-3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylurea(11 mg, 0.019 mmol), triethylamine (9.67 mg, 0.096 mmol), and(Z)-3-chloroacrylic acid (2.43 mg, 0.023 mmol) in DCM (0.4 ml, 6.21mmol) was added a solution of2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphinane 2,4,6-trioxide (21.9mg, 0.034 mmol) at 0° C., and the resulting mixture was stirred at roomtemperature for 30 min. The mixture was concentrated and the residue waspurified by silica gel column to obtain the title compound (7.5 mg,yield: 59%). ¹H NMR (400 MHz, CDCl₃) δ 1.29 (t, J=8.0 Hz, 3H) 2.90 (br.s., 4H) 3.27 (s, 3H) 3.45 (t, J=4.64 Hz, 4H) 3.91 (s, 6H) 5.90 (s, 1H)6.32 (d, J=8.41 Hz, 1H) 6.52 (s, 1H) 6.59 (d, J=8.41 Hz, 1H) 6.73 (dd,J=8.78, 2.76 Hz, 1H) 7.09 (br. s., 1H) 7.24 (s, 2H) 7.83 (br. s., 1H)8.36 (s, 1H) 8.69 (br. s., 1H) 12.52 (s, 1H); MS (ESI): 663.1 [M+H]⁺.

Example—185

N-(2-((6-(3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methylureido)pyrimidin-4-yl)amino)-6-fluorophenyl)acrylamide

The compound was synthesized following the approach outlined inProcedure 2G (Example 123), substituting 3-fluoro-2-nitroaniline in step(d), omitting step (e), and modifying step (g) to the followingprocedure: To a solution of1-(6-((2-amino-3-methylphenyl)amino)pyrimidin-4-yl)-3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-methyl-3-((2-(trimethylsilyl)ethoxy)methyl)urea(31 mg, 0.051 mmol), diisopropyl ethylamine (13.1 mg, 0.103 mmol), andacrylic acid (39.9 mg, 0.055 mmol) in DCM (1 ml, 15.54 mmol) was added asolution of 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphinane2,4,6-trioxide (48.1 mg, 0.076 mmol, 50% solution EtOAc) at 0° C., andthe resulting mixture was stirred at room temperature for 2 h. Themixture was concentrated and the residue was purified by silica gelcolumn to obtain the SEM protected title compound (24 mg, yield: 71% inthree steps). Following the final step (g), the title compound wasisolated (12 mg, yield: 62%). ¹H-NMR (400 MHz, CDCl₃) δ 3.40 (s, 3H)3.93 (m, 6H) 5.91 (d, 1H) 6.20 (s, 1H) 6.30-6.45 (m, 1H) 6.50-6.58 (m,2H) 7.01 (t, 1H) 7.28-7.35 (i, 1H) 7.42-7.63 (m, 2H) 8.41 (s, 1H) 12.43(br. s., 1H); ESI-MS: 535 [M+H]⁺.

Assays of Biological Activity

Assay of Binding to FGFR4.

Purified, recombinant FGFR4 was pre-incubated with 10 μM compoundovernight at 4° C., or for 1 hour at room temperature. Followingpre-incubation, protein samples were separated using SDS-PAGE and gelswere stained with SimplyBlue™ SafeStain (Life Technologies, GrandIsland, N.Y.). FGFR bands were cut out and digested using an In-GelTryptic Digestion Kit (Thermo Scientific, Waltham, Mass.). Digestedsamples were run on a Thermo Scientific Q Exactive™ LCMS using reversephase separation and tandem mass spectrometry to identify modifiedpeptides.

Alternatively, following pre-incubation FGFR4 was concentrated andbuffer exchanged on an OPTI-TRAP protein concentrating and desalting C4column (Optimize Technologies). Protein was eluted in acetonitrilecontaining 0.1% formic acid and run by direct injection on a ThermoScientific Q Exactive™ LCMS to identify modified, intact FGFR4.

Results provided below in Table 2 confirm covalent adduct formation ofthe tested compounds with the peptides by correspondence of the expectedmass of the peptide-ligand adduct with the mass observed.

TABLE 2 Expected mass Observed mass Compound # [Da] [Da] 100 3133.503134.48 100 35835.9 35836.1 108 35948.0 35948.1IC₅₀ Profiling of Kinase Activity Inhibition.

Compounds were profiled for FGFR inhibition activity at Reaction BiologyCorporation (Malvern, Pa.) with their Kinase HotSpot℠ assay. See,Anastassiadis et al., 2011, Comprehensive assay of kinase catalyticactivity reveals features of kinase inhibitor selectivity. NatBiotechnol 29, 1039-1045.

Recombinant FGFR1 (2.5 nM), FGFR2 (1 nM), FGFR3 (5 nM), or FGFR4 (12 nM)(Invitrogen™) was prepared as a mixture with substrate KKKSPGEYVNIEFG(SEQ ID NO:1) (20 μM, FGFR1 substrate); and Poly [E,Y] 4:1 (0.2 mg/ml,FGFR2,3,4 substrate)] in kinase reaction buffer (20 mM HEPES-HCl, pH7.5, 10 mM MgCl₂, 2 mM MnCl₂, 1 mM EGTA, 0.02% Brij35, 0.1 mM Na₃VO₄,0.02 mg/ml BSA, 2 mM DTT, and 1% DMSO). Compound was added to theenzyme/substrate mixture using acoustic technology (Labcyte® Echo 550,Sunnyvale, Calif.) (see, Olechno et al., 2006, Improving IC₅₀ resultswith acoustic droplet ejection. JALA 11, 240-246) and pre-incubated for0, 15, or 60 minutes at room temperature. After compound pre-incubation,a mixture of ATP (Sigma-Aldrich®) and ³³P-γ-ATP (PerkinElmer) was addedto a final concentration of 10 μM to initiate kinase reactions.Reactions were incubated for 120 minutes at room temperature and thenspotted onto Whatman™ P81 ion exchange filter paper. Unbound phosphatewas removed by extensively washing filters in 0.75% phosphoric acid.See, Anastassiadis et al., 2011, Comprehensive assay of kinase catalyticactivity reveals features of kinase inhibitor selectivity. NatBiotechnol 29, 1039-1045.

Results for FGFR4 and FGFR1 are shown next to individual compoundslisted in Table 1 above. The compounds showed selective inhibition ofFGFR4, with a higher IC₅₀ for FGFR1.

Without wishing to be bound by theory, the IC₅₀ activity with respect toFGFR1 is generally representative of the activity with respect to FGFR1,FGFR2, and FGFR3. See also, Dieci et al., 2013, Fibroblast Growth FactorReceptor Inhibitors as a Cancer Treatment: From a Biologic Rationale toMedical Perspectives. Cancer Discovery, F1-F16.

To confirm, some of the compounds were also tested for FGFR2 and FGFR3inhibition. These results shown below in Table 3 are consistent with theIC₅₀ activity of FGFR1 being generally representative of the activity ofFGFR1, FGFR2, and FGFR3, and further demonstrates the selectivity ofthese FGFR4 inhibitors.

TABLE 3 Compound FGFR2 FGFR3 FGFR1 FGFR4 # IC₅₀ (μM) IC₅₀ (μM) IC₅₀ (μM)IC₅₀ (μM) 100 4.18 1.98 >10.1 <0.001 108 1.98 2.00 0.173 <0.001In Vivo Efficacy in Tumor Models.

Compound 108 was evaluated for its ability to inhibit tumor growth innude mice bearing tumor xenografts from three different humanhepatocellular carcinoma tumor cell lines. These cell lines arerepresentative of cancers having an altered FGFR4 and/or FGF19 status.See Sawey et al., Cancer Cell 19(3): 347-358 (2011).

Animals:

Nude mice, ages 6-8 weeks, and weighing approximately 19-25 g, werepurchased from Taconic (Taconic, Hudson, N.Y.). All animal experimentswere done in accordance with protocols approved by the InstitutionalAnimal Care and Use Committee.

Tumor Xenografts and Treatment:

7.5×10⁶ HUH7 cells (HSRRB cat. no. JCRB0403), 5×10⁶ Hep3B (ATCC cat. no.HB8064), or 2.5×10⁶ JHH7 cells (HSRRB cat. no. JCRB1031), each in atotal volume of 100 μl, 1:1 Matrigel (Corning Inc, Corning, N.Y.), wereinjected subcutaneously (s.c.) into the right lateral flank. When tumorsreached 150-200 mm³, the mice were randomized into treatment groups of5-10 animals. Dosing was performed twice daily by intraperitonealinjection at the indicated dosages for 15 days using Compound 108,formulated in a vehicle of 5% DMSO (Alfa Aesar, Ward Hill, Mass.), 10%PEG300 (Sigma, St. Louis, Mo.), 8% TWEEN® 80 (Sigma, St. Louis, Mo.),77% USP Saline at the desired concentration. Tumor volumes werecollected twice weekly using the formula Volume=(length*width²)/2. Bodyweights were collected twice weekly, as well. All animals were observedand cared for in accordance with The Guide for Care and Use ofLaboratory Animals, 8th edition (National Academies Press, WashingtonD.C.).

Statistical Methods:

Statistical comparisons were made at the end of the experiment usingRepeated Measures Anova with Bonferroni post-test for comparisons oftreatment groups, using GraphPad Prism 5. The following criteria wereused to determine Progressive Disease, Stable Disease, PartialRegression, and Complete Regression. Progressive Disease is defined asthree consecutive measurements increasing from best response or >120%initial tumor volume. Stable Disease is three consecutive measurements<120% and >50% of initial tumor volume, whereas three consecutivemeasurements <50% initial tumor volume qualifies as a PartialRegression. A Complete Regression is three consecutive measurements <30mm³. Chi-squared test was used to compare responses between treatmentgroups (Microsoft Excel).

Results from animals bearing tumors from HUH7, HEP3B, and JHH7 cancercells are shown in FIGS. 1-3, respectively, and are also reflected inTable 4.

TABLE 4 Inhibition of Tumor Growth in FGF19 amplified HCC xenograftsDose Complete Partial Stable Progressive (mg/kg) Regression RegressionDisease Disease HUH7 (n = 10 per group) 25 1 4 3  2 37.5 2 5 3  0 HEP3B(n = 5 per group) 12.5 0 0 0  5 25 0 1 4  0 37.5 5 0 0  0 JHH7 (n = 10per group) 12.5 0 0 0 10 25 0 0 0 10 37.5 0 0 0 10

These data demonstrate that compound 108 is efficacious in all models.Among the three models, HEP3B is the most sensitive, JHH7 the leastsensitive and HUH7 showing intermediate sensitivity to compound 108.Although a dose response can be seen in FIG. 3 for JHH7, there wasProgressive Disease in all dose levels tested.

Comparative Studies of Compound 108 with BGJ398.

Comparative studies were done with Compound 108 and the known FGFRinhibitor BJG398.

Biochemical Kinase Assay Protocol to Obtain IC₅₀:

Recombinant FGFR1 (2.5 nM), or FGFR4 (12 nM) was prepared as a mixturewith substrate KKKSPGEYVNIEFG (SEQ ID NO:1) (20 μM, FGFR1 substrate);Poly [E,Y] 4:1 (0.2 mg/ml, FGFR2,3,4 substrate)] in kinase reactionbuffer (20 mM HEPES-HCl, pH 7.5, 10 mM MgCl₂, 2 mM MnCl₂, 1 mM EGTA,0.02% Brij35, 0.1 mM Na3VO4, 0.02 mg/ml BSA, 2 mM DTT, and 1% DMSO).Compound was added to the enzyme/substrate mixture using acoustictechnology and pre-incubated for 0, 15, or 60 minutes at roomtemperature. After compound pre-incubation, ³³P-γ-ATP was added at afinal concentration of 10 μM to initiate kinase reactions. Reactionswere incubated for 120 minutes at room temperature. Substratephosphorylation was monitored by filter assay, as above. Results areshown in Table 5. The results reported show that compound 108 is a morepotent FGFR4 inhibitor, whereas BGJ398 is a more potent FGFR1 inhibitor.

TABLE 5 Comparative Testing of Compound 108 and BGJ398 with BiochemicalKinase assay Compound 108 BGJ398 Kinase IC₅₀ (nM) IC₅₀ (nM) FGFR4 <0.213 FGFR1 513 1.0Cellular Viability Assay Protocol to Obtain GI₅₀:

Cells lines were cultured at 37° C., 5% CO₂ and 95% humidity. Culturemedia were purchased from GIBCO®, USA. For viability assay, 2000cells/well were seeded in 96 well plates, incubated for 24 h beforecompound treatment. Following compound addition, plates were incubatefor 72 h at 37° C. with 5% CO₂, and then measured by means of CTG assay(CellTiter-Glo® Luminescent Cell Viability Assay, Cat. No.: G7572,Promega). Results are shown in Table 6. The table shows compound 108 ismore potent than BGJ398 in Hep3B cells, an FGF19 amplified line. Thepotency in HUH7 and JHH7, the other two FGF19 amplified lines, arecomparable between compound 108 and BGJ398. HepG2 (ATCC cat. no.HB-8065), SNU398 (ATCC cat. no. CRL-2233) and SNU449 (ATCC cat. no.CRL-2234) are FGF19 non-amplified cell lines that were used as controls.

GI₅₀ is the concentration of test drug where 100×(T−T0)/(C−T0)=50. See,e.g., Monks et al., Feasibility of a High-Flux Anticancer Drug ScreenUsing a Diverse Panel of Cultured Human Tumor Cell Lines, J Natl CancerInst (1991) 83(11):757-766; Boyd et al., Data Display and AnalysisStrategies for the NCI Disease-oriented In Vitro Antitumor Drug Screen,in CYTOTOXIC ANTICANCER DRUGS: MODELS AND CONCEPTS FOR DRUG DISCOVERYAND DEVELOPMENT, Valeriote et al., eds. (1990), pp. 11-34. Theluminescence of the test well after a 72 h period of exposure to testdrug is T, the luminescence at time zero is T0, and the controlluminescence is C. The GI₅₀ measures the growth inhibitory power of thetest agent.

TABLE 6 Comparative Testing of Compound 108 and BGJ398 in CellularViability assays Compound 108 BGJ398 Cell Line GI₅₀ (nM) GI₅₀ (nM) HEP3B18 ± 6 nM 74 ± 23 nM (n = 27) (n = 6) JHH7 216 ± 70 nM 178 ± 30 nM (n =4) (n = 2) HUH7 408 ± 128 nM 231 ± 100 nM (n = 4) (n = 2) HEPG2 6506 ±1424 nM 2260 ± 1182 nM (n = 27) (n = 6) SNU398 >10,000 (n = 2) notmeasured SNU449 >10,000 (n = 2) not measuredIn Vivo Efficacy Comparison:

Nude mice were used for these experiments as above. 5.0×10⁶ Hep3B cellsin a total volume of 100 μl, 1:1 Matrigel (Corning Inc, Corning, N.Y.),were injected s.c. into the right lateral flank. When tumors reached150-200 mm³ the mice were randomized into treatment groups of 5-10animals. Treatment was then started using Compound 108, formulated in avehicle of 5% DMSO (Alfa Aesar, Ward Hill, Mass.), 10% PEG300 (Sigma,St. Louis, Mo.), 8% TWEEN® 80 (Sigma, St. Louis, Mo.), 77% USP Saline atthe desired concentration. BGJ398, formulated as a suspension in 0.5%Methylcellulose (Sigma)/0.2% TWEEN® 80, was suspended at the desiredconcentration. Both drugs were dosed for 18 dyas, except for onetreatment group (see below). Tumor volumes were collected twice weeklyusing the formula Volume=(length*width²)/2. Body weights were collectedtwice weekly as well. All animals were observed and cared for inaccordance with The Guide for Care and Use of Laboratory Animals, 8thedition (National Academies Press, Washington D.C.). The results of thiscomparative in vivo study are shown in FIG. 4.

The data show that compound 108 is more efficacious than BGJ398 attolerable dosage levels. Although BGJ398 at 60 mg/kg showed efficacycomparable to compound 108, the dosing of this BGJ398 60 mg/kg group hadto be terminated on Day 11 due to poor health of animals. Thisdifference in toxicity is not due to routes of administration becausethe group of animals dosed orally with BGJ398 at 30 mg/kg did notexhibit poor health.

The foregoing is illustrative of the present invention, and is not to beconstrued as limiting thereof. The invention is defined by the followingclaims, with equivalents of the claims to be included therein.

We claim:
 1. A method of treating sarcoma in a subject in need thereofcomprising administering to said subject a treatment effective amount ofa compound of Formula I:

wherein: R³ is selected from the group consisting of: C₁₋₆alkyl,C₁₋₆alkoxyC₁₋₆alkyl, NR¹⁰R¹¹C₁₋₆alkyl, R¹⁰heterocyclylC₁₋₆alkyl,R¹⁰arylC₁₋₆alkyl, and R¹⁰heteroarylC₁₋₆alkyl, wherein R¹⁰ and R¹¹ areeach independently selected from the group consisting of: hydrogen andC₁₋₆alkyl; E is selected from the group consisting of:—NR¹³C(O)CR¹⁴═CHR¹⁵, and —NR¹³C(O)C≡CR¹⁴, wherein R¹³ is selected fromthe group consisting of: hydrogen and methyl, and R¹⁴ and R¹⁵ are eachindependently selected from the group consisting of: hydrogen, methyl,fluoro and chloro; R¹² is selected from the group consisting of:hydrogen, halo, C₁₋₆alkyl, C₁₋₆alkoxy, hydroxyC₁₋₆alkyl,hydroxyC₁₋₆alkoxy, C₁₋₆alkoxyC₁₋₆alkoxy, C₁₋₆alkoxyC₁₋₆alkyl,R⁵R⁶heterocyclyl, —C(O)heterocyclylR⁵R⁶, R⁵R⁶heterocyclylC₁₋₆alkyl,NR⁵R⁶, NR⁵R⁶C₁₋₆alkyl, —C(O)NR⁵R⁶, and NR⁵R⁶C₁₋₆alkoxy, wherein R⁵ andR⁶ are each independently selected from the group consisting ofhydrogen, C₁₋₆alkyl, hydroxyC₁₋₆alkyl, aminoC₁₋₆alkyl, —C(O)C₁₋₆alkyland C₁₋₆alkylsulfonyl; and R¹ is phenyl, wherein said phenyl issubstituted 2, 3, or 4 times with independently selected halo orC₁₋₆alkoxy, or a pharmaceutically acceptable salt thereof.
 2. The methodof claim 1, wherein R³ is selected from the group consisting of: methyl,methoxyethyl, 4-pyridylmethyl, 3-pyridylmethyl, 2-pyridylmethyl, benzyl,N,N-dimethylaminopropyl, 3-methylisoxazol-5-yl-methyl, and4-methylpiperazin-1-yl-propyl.
 3. The method of claim 1, wherein E is—NHC(O)CH═CH₂.
 4. The method of claim 1, wherein R¹² is selected fromthe group consisting of: hydrogen, fluoro, chloro, methyl, methoxy,N,N-dimethylaminoethyl, piperazin-1-yl, 4-ethylpiperazin-1-yl,4-ethylpiperazin-1-yl-methyl, 1-methylpiperidine-4-yl,1-ethylpiperidine-4-yl, N,N-dimethylaminomethyl,N,N-dimethylaminopropyl, piperidine-4-yl, morpholino,3,5-dimethylpiperazin-1-yl, 4-(methylsulfonyl)piperazin-1-yl,N,N-dimethylaminoethoxy, 4-(2-hydroxyethyl)piperazin-1-yl,hydroxyethoxy, methoxyethoxy, hydroxymethyl, methoxymethyl,2-methoxypropyl, 2-hydroxypropyl, 2-aminopropyl,4-methylpiperazin-1-yl-carbonyl, 4-ethylpiperazin-1-yl-carbonyl,4-[2-propyl]piperazin-1-yl, 4-acetylpiperazin-1-yl,N-methyl-N-hydroxyethyl-amino, N,N-dimethylamido, and4-(2-aminoethyl)piperazin-1-yl.
 5. The method of claim 1, wherein R¹² isR⁵R⁶heterocyclyl.
 6. The method of claim 1, wherein R¹² is4-ethylpiperazin-1-yl.
 7. The method of claim 1, wherein R¹ is2,6-dichloro-3,5-dimethoxyphenyl.
 8. The method of claim 1, wherein saidcompound or salt is a compound of Formula I(a):

or a pharmaceutically acceptable salt thereof.
 9. The method of claim 1,wherein said compound is selected from the group consisting of:

or a pharmaceutically acceptable salt thereof.
 10. The method of claim1, wherein the compound is:

or a pharmaceutically acceptable salt thereof.
 11. The method of claim1, wherein the compound is


12. The method of claim 1, wherein said sarcoma is rhabdomyosarcoma. 13.The method of claim 12, wherein said rhabdomyosarcoma has FGFR₄expression.
 14. The method of claim 12, wherein said rhabdomyosarcomahas FGFR₄ expression.
 15. The method of claim 10, wherein said sarcomais rhabdomyosarcoma.
 16. The method of claim 15, wherein saidrhabdomyosarcoma has FGFR₄ expression.
 17. The method of claim 15,wherein said rhabdomyosarcoma has FGFR₄ expression.
 18. The method ofclaim 11, wherein said sarcoma is rhabdomyosarcoma.
 19. The method ofclaim 18, wherein said rhabdomyosarcoma has FGFR₄ expression.
 20. Themethod of claim 18, wherein said rhabdomyosarcoma has FGFR₄ expression.